Showing papers in "Genetics in 1984"

Genes affecting the regulation of suc2 gene expression by glucose repression in saccharomyces cerevisiae

Abstract: Mutants of Saccharomyces cerevisiae with defects in sucrose or raffinose fermentation were isolated. In addition to mutations in the SUC2 structural gene for invertase, we recovered 18 recessive mutations that affected the regulation of invertase synthesis by glucose repression. These mutations included five new snf1 (sucrose nonfermenting) alleles and also defined five new complementation groups, designated snf2, snf3, snf4, snf5 and snf6. The snf2, snf4 and snf5 mutants produced little or no secreted invertase under derepressing conditions and were pleiotropically defective in galactose and glycerol utilization, which are both regulated by glucose repression. The snf6 mutant produced low levels of secreted invertase under derepressing conditions, and no pleiotropy was detected. The snf3 mutants derepressed secreted invertase to 10–35% the wild-type level but grew less well on sucrose than expected from their invertase activity; in addition, snf3 mutants synthesized some invertase under glucose-repressing conditions.—We examined the interactions between the different snf mutations and ssn6, a mutation causing constitutive (glucose-insensitive) high-level invertase synthesis that was previously isolated as a suppressor of snf1 . The ssn6 mutation completely suppressed the defects in derepression of invertase conferred by snf1, snf3, snf4 and snf6, and each double mutant showed the constitutivity for invertase typical of ssn6 single mutants. In contrast, snf2 ssn6 and snf5 ssn6 strains produced only moderate levels of invertase under derepressing conditions and very low levels under repressing conditions. These findings suggest roles for the SNF1 through SNF6 and SSN6 genes in the regulation of SUC2 gene expression by glucose repression.

Mutants of S. cerevisiae defective in the maintenance of minichromosomes.

Abstract: We have isolated yeast mutants that are defective in the maintenance of circular minichromosomes. The minichromosomes are mitotically stable plasmids, each of which contains a different ARS (autonomously replicating sequence), a centrometeric sequence, CEN5 , and two yeast genes, LEU2 and URA3 . Forty minichromosome maintenance-defective (Mcm - ) mutants were characterized. They constitute 16 complementation groups. These mutants can be divided into two classes, specific and nonspecific, by their differential ability to maintain minichromosomes with different ARS s. The specific class of mutants is defective only in the maintenance of minichromosomes that carry a particular group of ARS s irrespective of the centromeric sequence present. The nonspecific class of mutants is defective in the maintenance of all minichromosomes tested irrespective of the ARS or centromeric sequence present. The specific class may include mutants that do not initiate DNA replication effectively at specific ARS s present on the minichromosomes; the nonspecific class may include mutants that are affected in the segregation and/or replication of circular plasmids in general.

Organization and mapping of a sequence on the Drosophila melanogaster X and Y chromosomes that is transcribed during spermatogenesis.

Abstract: The D. melanogaster DNA segment in the recombinant phage λDm2L1 contains at least eight copies of a tandemly repeated 1250-base pair (bp) sequence (henceforth called the 2L1 sequence). Testes from XO D. melanogaster males contain an abundant 800-base RNA species that is homologous to a 520-bp region of the 2L1 sequence. Blotting experiments show that the 2L1 sequence is repeated in the D. melanogaster genome and is present on both the X and Y chromosomes. With the use of X-Y translocations, the 2L1 sequence has been mapped to a region between kl-1 and kl-2 on the long arm of the Y chromosome. In Oregon-R wild type there are an estimated 200 copies of the 2L1 sequence on the X chromosome and probably at least 80 copies on the Y chromosome. In some other strains the repetition frequency on the Y chromosome is about the same, but the copy number on the X chromosome is much reduced. On the basis of the five strains investigated, there is a correlation between copy number of the 2L1 sequence on the X chromosome and the presence of a particular allele of the Stellate locus (Ste; 1-45.7). It seems that low copy number corresponds to Ste + and high copy number corresponds to Ste. The Ste locus determines whether single or star-shaped crystals are observed in the spermatocytes of XO males. Studies using D. simulans and D. mauritiana DNA show that the 2L1 sequence is homologous to restriction fragments in male DNA but not female DNA, indicating that this sequence is present only on the Y chromosome in these two species. In DNA derived from D. erecta, D. teissieri and D. yakuba, there is very little, if any, hybridization with the 2L1 sequence probe.

Polymorphic sites and the mechanism of evolution in human mitochondrial dna

Abstract: Twelve restriction enzymes were used to screen for the presence or absence of cleavage sites at 441 locations in the mitochondrial DNA of 112 humans from four continents. Cleavage maps were constructed by comparison of DNA fragment sizes with those expected from the published sequence for one human mtDNA. One hundred and sixty-three of the sites were polymorphic, i.e., present in some individuals but absent from others, 278 sites being invariant. These polymorphisms probably result from single base substitutions and occur in all functional regions of the genome.—In 77 cases, it was possible to specify the exact nature and location (within a restriction site) of the mutation responsible for the absence of a restriction site in a known human mtDNA sequence and its presence in another human mtDNA. Fifty-two of these 77 gain mutations occur in genes coding for proteins, 34 being silent and 18 causing amino acid replacements; moreover, nine of the replacements are radical.—Notable also is the anomalous ratio of transitions to transversions required to account for these 77 restriction site differences between the known human mtDNA sequences and other human mtDNAs. This ratio is lower for most groups of restriction sites than has been reported from sequence comparisons of limited parts of the mtDNA genome in closely related mammals, perhaps indicating a special functional role or sensitivity to mutagenesis for palindromic regions containing high levels of guanine and cytosine.—From the genomic distribution of the 163 polymorphic sites, it is inferred that the level of point mutational variability in tRNA and rRNA genes is nearly as high as in protein-coding genes but lower than in noncoding mtDNA. Thus, the functional constraints operating on components of the protein-synthetic apparatus may be lower for mitochondria than for other systems. Furthermore, the mitochondrial genes for tRNAs that recognize four codons are more variable than those recognizing only two codons.—Among the more variable of the human mitochondrial genes coding for proteins is that for subunit 2 of cytochrome oxidase; this polypeptide appears to have been evolving about five times faster in primates than in other mammals. Cytochrome c, a nuclearly encoded protein that interacts directly with the oxidase 2 subunit in electron transport, has also evolved faster in primates than in rodents or ungulates. This example, along with that for the mitochondrial rRNA genes and the nuclear genes coding for mitochondrial ribosomal proteins, provides evidence for coevolution between specific nuclear and mitochondrial genes.

Mutations affecting Ty-mediated expression of the HIS4 gene of Saccharomyces cerevisiae.

Abstract: We have identified mutations in seven unlinked genes ( SPT genes) that affect the phenotypes of Ty and δ insertion mutations in the 5′ noncoding region of the HIS4 gene of S. cerevisiae. Spt mutants were selected for suppression of his4-912δ , a solo δ derivative of Ty912. Other Ty and δ insertions at HIS4 are suppressed by mutations in some but not all of the SPT genes. Only spt4 suppresses a non-Ty insertion at HIS4 . In addition to their effects on Ty and δ insertions, mutations in several SPT genes show defects in general cellular functions—mating. DNA repair and growth.

Ploidy barrier to endosperm development in maize

Abstract: Maize kernels inheriting the indeterminate gametophyte mutant (ig) on the female side had endosperms that ranged in ploidy level from diploid (2x) to nonaploid (9x). In crosses with diploid males, only kernels of the triploid endosperm class developed normally. Kernels of the tetraploid endosperm class were half-sized but with well-developed embryos that regularly germinated. Kernels of endosperm composition other than triploid or tetraploid were abortive.-Endosperm ploidy level resulting from mating ig/ig x tetraploid Ig similarly was variable. Most endosperms started to degenerate soon after pollination and remained in an arrested state. Hexaploid endosperm was exceptional; it developed normally during the sequence of stages studied and accounted for plump kernels on mature ears. Since such kernels have diploid maternal tissues (pericarp) but triploid embryos, the present finding favors the view that endosperm failure or success in such circumstances is governed by conditions within the endosperm itself.-Whereas tetraploid endosperm consisting of three maternal genomes and one paternal genome is slightly reduced in size but supports viable seed development, that endosperm having two maternal and two paternal chromosome sets was highly defective and conditioned abortion. Thus, development of maize endosperm evidently is affected by the parental source of its sets of chromosomes.

Autoregulatory functioning of a drosophila gene product that establishes and maintains the sexually determined state

Abstract: Sxl appears to head a regulatory gene hierarchy that controls Drosophila sexual dimorphism in response to the X chromosome/autosome balance. Only XXAA cells normally have Sxl(+) activity. It maintains both the female morphogenetic sequence and a level of X-linked dosage-compensated gene expression compatible with diplo-X cell survival. In the absence of this activity, male sexual development and dosage-compensated gene hyperactivation ensure. Loss-of-function Sxl mutations generally have female-specific lethal effects caused by upsets in dosage compensation. New female-viable Sxl mutant alleles and combinations which lack Sxl's female sex determination function, yet still provide sufficient dosage compensation function for diplo-X survival, are described here. Consequently, such mutants cause genotypic females to develop as phenotypic males. Some of these sex-transforming Sxl mutants do not require the maternally produced da(+) activity that is normally essential for the functioning of zygotic Sxl alleles. In this paper, products of these unusual alleles are shown to act in trans to induce the expression of zygotic Sxl(+) alleles that would otherwise be unable to function due to a lack of maternal da(+) activity. This result indicates a third function for Sxl(+) product: a positive autoregulatory role. Controls for the autoregulation experiments demonstrated the sex-trans-forming epigenetic effect of the da mutation for the first time in diploids. In these experiments the female-specific zygotic lethal effects that normally would have accompanied loss of maternal da(+) activity were suppressed by mutations known to block dosage-compensation gene hyperactivation-the autosomal, male-specific lethals. Three types of abnormal sexual phenotypes were produced in the experiments described here, each with important implications for the mechanism of sex determination: (1) a true intersex phenotype produced by one particular Sxl allele shows that Sxl(+) must be involved in the cellular response to the X/A balance rather than in its establishment; (2) a maternally induced, female-sterile phenotype indicates that either the process of autoregulation or the mutants used to demonstrate it are tissue specific and (3) a mosaic intersexual phenotype whose character implies that the Sxl(+ ) activity level is set early in development, both by the da( +)-mediated X/A balance signal and by autoregulation, and is maintained subsequently in a cell autonomous fashion, independent of the initiating X/A balance signal. Thus, this study supports the view that sex determination is truly determinative in the standard developmental sense, and that Sxl is the carrier of the sexually determined state.

A chromosome replication pattern deduced from pericarp phenotypes resulting from movements of the transposable element, modulator, in maize

Abstract: Modulator (Mp) was mapped after it transposed from the P locus on chromosome 1 by studying 105 light variegated/red twin sectors on medium variegated pericarp ears. Sixty-one percent of the receptor sites were detectably linked to P, and these showed an asymmetry of distribution adjacent to P. No transpositions were mapped in the 4 map units proximal to P, whereas 23 cases mapped to the same length distal to P. The remaining transpositions of Mp on chromosome 1, both proximal and distal to P, were equally scattered. It has previously been shown that when Modulator transposes it replicates at the P locus and a second time at the receptor site. The pattern of transposition adjacent to P is consistent with a hypothesis that a replicon initiation site is situated proximal to P; that Modulator transposes at the time of replication; that it is not able to transpose into a replicated region but only into a replicating one. No difference in distribution of receptor sites was found when the Modulator was detected vs. not detected in the red co-twins by testing with a Dissociation element.

The nature of nucleotide sequence divergence between barley and maize chloroplast dna

Abstract: Analysis of a 2175-base pair (bp) Sma I- Hin dIII fragment of barley chloroplast DNA revealed that rbcL (the gene for the large subunit of ribulose 1,5-bisphosphate carboxylase) and atpB (the gene for the β subunit of ATPase) are transcribed divergently and are separated by an untranscribed region of 155-166 bp. The rbcL mRNA has a 320-residue untranslated leader region, whereas the atpB mRNA has a 296- to 309-residue leader region. The sequence of these regions, together with the initial 113 bp of the atpB -coding region and the initial 1279 bp of the rbcL -coding region, is compared with the analogous maize chloroplast DNA sequences. Two classes of nucleotide differences are present, substitutions and insertions/deletions. Nucleotide substitutions show a 1.9-fold bias toward transitions in the rbcL -coding region and a 1.5-fold bias toward transitions in the noncoding region. The level of nucleotide substitutions between the barley and maize sequences is about 0.065/bp. Seventy-one percent of the substitutions in the rbcL -coding region are at the third codon position, and 95% of these are synonymous changes. Insertion/deletion events, which are confined to the noncoding regions, are not randomly distributed in these regions and are often associated with short repeated sequences. The extent of change for the noncoding regions (about 0.093 events/bp) is less than the extent of change at the third codon positions in the rbcL -coding region (about 0.135 events/bp), including insertion/delection events. Limited sequence analysis of the analogous DNA from a wild line ( Hordeum spontaneum ) and a primitive Iranian barley ( H. vulgare ) suggested a low rate of chloroplast DNA evolution. Compared to spinach chloroplast DNA, the barley rbcL-atpB untranslated region is extremely diverged, with only the putative rbcL promoters and ribosome-binding site being extensively conserved.

Hybridization and introgression among species of sunfish (lepomis): analysis by mitochondrial dna and allozyme markers

Abstract: We explore the potential of mitochondrial DNA (mtDNA) analysis, alone and in conjunction with allozymes, to study low-frequency hybridization and introgression phenomena in natural populations. MtDNAs from small samples of nine species of sunfish (Lepomis, Centrarchidae) were purified and digested with each of 13 informative restriction enzymes. Digestion profiles for all species were highly distinct: estimates of overall fragment homology between pairs of species ranged from 0–36%. Allozymes encoded by nine nuclear genes also showed large frequency differences among species and together with mtDNA provided many genetic markers for hybrid identification. A genetic analysis of 277 sunfish from two locations in north Georgia revealed the following: (1) a low frequency of interspecific hybrids, all of which appeared to be F1's; (2) the involvement of five sympatric Lepomis species in the production of these hybrids; (3) no evidence for introgression between species in our study locales (although for rare hybridization, most later-generation backcrosses would not be reliably distinguished from parentals); (4) a tendency for hybridizations to take place preferentially between parental species differing greatly in abundance; (5) a tendency for the rare species in a hybrid cross to provide the female parent. Our data suggest that absence of conspecific pairing partners and mating stimuli for females of rarer species may be important factors in increasing the likelihood of interspecific hybridization. The maternal inheritance of mtDNA offers at least two novel advantages for hybridization analysis: (1) an opportunity to determine direction in hybrid crosses; and (2) due to the linkage among mtDNA markers, an increased potential to distinguish effects of introgression from symplesiomorphy or character convergence.

Deviations from hardy-weinberg proportions: sampling variances and use in estimation of inbreeding coefficients

Abstract: An analysis is made of the distribution of deviations from Hardy-Weinberg proportions with k alleles and of estimates of inbreeding coefficients (f) obtained from these deviations. If f is small, the best estimate of f in large samples is shown to be 2 sigma i(Tii/Ni)/(k - 1), where Tii is an unbiased measure of the excess of the ith homozygote and Ni the number of the ith allele in the sample [frequency = Ni/(2N)]. No extra information is obtained from the Tij, where these are departures of numbers of heterozygotes from expectation. Alternatively, the best estimator can be computed from the Tij, ignoring the Tii. Also (1) the variance of the estimate of f equals 1/(N(k - 1] when all individuals in the sample are unrelated, and the test for f = 0 with 1 d.f. is given by the ratio of the estimate to its standard error; (2) the variance is reduced if some alleles are rare; and (3) if the sample consists of full-sib families of size n, the variance is increased by a proportion (n - 1)/4 but is not increased by a half-sib relationship. If f is not small, the structure of the population is of critical importance. (1) If the inbreeding is due to a proportion of inbred matings in an otherwise random-breeding population, f as determined from homozygote excess is the same for all genes and expressions are given for its sampling variance. (2) If the homozygote excess is due to population admixture, f is not the same for all genes. The above estimator is probably close to the best for all f values.

Measuring Gene Flow among Populations Having High Levels of Genetic Fragmentation

Abstract: We present an analysis of the genetic structures of 22 species of salamanders, with regard to levels of gene flow among populations. We estimate the gene flow parameter, Nm (the product of the effective population number and rate of migration among populations) using two alternative methods described by Wright and Slatkin. For most species, these two methods give approximately congruent estimates of Nm; when estimates differ, the method of Wright produces values slightly larger than those derived by the method of Slatkin. We analyze these results in light of independently derived historical inferences of the fragmentation of populations. This analysis suggests that the Nm values calculated from protein polymorphisms may contain information more relevant to historical patterns of gene exchange than to the current population dynamics; moderately large values of Nm may be calculated for species containing populations known to be no longer exchanging genes. Application of a method for estimating the maximum possible rate of gene exchange among populations indicates that, for most species studied here, gene flow among populations probably is no greater than the mutation rate. We suggest that most plethodontid species cannot be viewed as units whose cohesion is maintained by continuing gene exchange. Furthermore, we suggest that phenotypic uniformity among populations is not easily explained by hypotheses of continual stabilizing selection and propose that future work concentrate upon clarification of the genetic and epigenetic factors conferring self-maintenance or autopoietic properties on living systems.

A Suppressor of snf1 Mutations Causes Constitutive High-Level Invertase Synthesis in Yeast

Abstract: The SNF1 gene product of Saccharomyces cerevisiae is required to derepress expression of many glucose-repressible genes, including the SUC2 structural gene for invertase. Strains carrying a recessive snf1 mutation are unable to ferment sucrose. We have isolated 30 partial phenotypic revertants of a snf1 mutant that were able to ferment sucrose. Genetic characterization of these revertants showed that the suppressor mutations were all recessive and defined eight complementation groups, designated ssn1 through ssn8 (suppressor of snf1 ). The revertants were assayed for secreted invertase activity, and although activity was detected in members of each complementation group, only the ssn6 strains contained wild-type levels. Synthesis of secreted invertase in ssn6 strains was found to be constitutive, that is, insensitive to glucose repression; moreover, the ssn6 mutations also conferred constitutivity in a wild-type ( SNF1 ) genetic background and are, therefore, not merely suppressors of snf1 . Pleiotropic defects were observed in ssn6 mutants. Genetic analysis suggested that the ssn6 mutations are allelic to the cyc8 mutation isolated by R. J. Rothstein and F. Sherman, which causes increased production of iso-2-cytochrome c. The data suggest a regulatory function for SSN6 .

A genetic analysis of targeted growth in mice

Abstract: Effects of normal growth regulation on components of phenotypic variance and covariance of body weight were examined in a cross-fostering study of growth between 2 and 10 wk of age in ICR randombred mice. Different early growth rates caused genetic, postnatal maternal and residual environmental variances to increase, but these variances were subsequently reduced by negative autocorrelation between early and later growth. Postnatal maternal variance continued to increase for about 1 wk after weaning but then decreased substantially. Genetic variance caused by preweaning growth followed a pattern of increase and decrease very similar to that of postnatal maternal variance, but this pattern was masked by new genetic variance. Normal growth regulation affects the magnitudes of genetic variances and serial autocorrelations. The timing of these changes suggests that regulation of cell numbers reduces variance near the end of exponential growth, but this may be obscured by subsequent increase in cell size. In contrast with earlier studies, we find that targeted growth reduces both genetically and environmentally determined differences among early growth trajectories. Final size may be determined by an antagonistic balance between early growth rate and age at initiation of puberty.

Identification of new genes involved in the regulation of yeast alcohol dehydrogenase II.

Abstract: Recessive mutations in two negative control elements, CRE1 and CRE2, have been obtained that allow the glucose-repressible alcohol dehydrogenase (ADHII) of yeast to escape repression by glucose. Both the cre1 and cre2 alleles affected ADHII synthesis irrespective of the allele of the positive effector, ADR1. However, for complete derepression of ADHII synthesis, a wild-type ADR1 gene was required. Neither the cre1 nor cre2 alleles affected the expression of several other glucose-repressible enzymes. A third locus, CCR4, was identified by recessive mutations that suppressed the cre1 and cre2 phenotypes. The ccr4 allele blocked the derepression of ADHII and several other glucose-repressible enzymes, indicating that the CCR4 gene is a positive control element. The ccr4 allele had no effect on the repression of ADHII when it was combined with the ADR1-5c allele, whereas the phenotypically similar ccr1 allele, which partially suppresses ADR1-5c, did not suppress the cre1 or cre2 phenotype. Complementation studies also indicated that ccr1 and snf1 are allelic. A model of ADHII regulation is proposed in which both ADR1 and CCR4 are required for ADHII expression. CRE1 and CRE2 negatively control CCR4, whereas CCR1 is required for ADR1 function.

Primary trisomics of rice: origin, morphology, cytology and use in linkage mapping

Abstract: Twelve primary trisomics of Oryza sativa L. were isolated from the progenies of spontaneous triploids and were transferred by backcrossing to the genetic background of IR36, a widely grown high yielding rice variety. Eleven trisomics can be identified morphologically from one another and from diploids. However, triplo 11 is difficult to distinguish from diploid sibs.-The extra chromosome of each trisomic was identified cytologically at pachytene stage of meiosis, and the chromosomes were numbered according to their length at this stage. The major distinguishing features of each pachytene chromosome were redescribed.-The female transmission rates varied from 15.5% for triplo 1, the longest chromosome, to 43.9% for triplo 12, the shortest chromosome. Seven of the 12 primary trisomics transmitted the extra chromosome through the male. The low level of chromosomal imbalance tolerated by rice and other evidence are interpreted to indicate that this species is a basic diploid.-Genetic segregation for 22 marker genes in the trisomic progenies was studied. Of a possible 264 combinations, involving 22 genes and 12 trisomics, 120 were examined. Marker genes for each of the 12 chromosomes were identified. The results helped establish associations between linkage groups and cytologically identifiable chromosomes of rice for the first time. Relationships between various systems of numbering chromosomes, trisomics, linkage groups and marker genes are described, and a revised linkage map of rice is presented.

Polyandry in honey bees (apis mellifera l.): sperm utilization and intracolony genetic relationships

Abstract: Sperm usage by queen honey bees was examined by progeny analyses using six phenotypically distinct genetic markers. No evidence was found for sperm displacement or precedence. All queens used the sperm of all males that inseminated them during all sampling periods. Sperm usage, as measured by phenotypic frequencies, did fluctuate nonrandomly but did not result in abnormally high representation of a single phenotype or the elimination of other phenotypes as has often been suggested. The genetic relationships of workers within honey bee colonies are estimated from the data presented. Average genetic relatedness is shown to be low among colony nestmates and probably approaches 0.25 in colonies with naturally mated queens. There is no evidence for elevated relatedness among colony subfamilies due to nonrandom fluctuations in sperm usage by queens or for numerical dominance of any subfamilies.

Population bottlenecks and nonequilibrium models in population genetics. i. allele numbers when populations evolve from zero variability

Abstract: A simple numerical method was developed for the mean number and average age of alleles in a population that was initiated with no genetic variation following a sudden population expansion. The methods are used to examine the question of whether allele numbers are elevated compared with values seen in equilibrium populations having equivalent gene diversity. Excess allele numbers in expanding populations were found to be the rule. This was true whether the population began with zero variation or with low levels of variation in either of two initial distributions (initially an equilibrium allele frequency distribution or initially with loci occurring in only two classes of variation). Although the increase of alleles may persist for only a short time, when compared with the time which is required for approach to final equilibrium, the increase may be long when measured in absolute generation numbers. The pattern of increase in very rare alleles (those present only once in a sample) and the persistence of the original allele were also investigated.

Formation of chromosome rearrangements by P factors in Drosophila.

Abstract: We studied a collection of 746 chromosome rearrangements all induced by the activity of members of the P family of transposable elements in Drosophila melanogaster. The chromosomes ranged from simple inversions to complex rearrangements. The distribution of complex rearrangement classes was of the kind expected if each rearrangement came about from a single multibreak event followed by random rejoining of chromosome segments, as opposed to a series of two-break events. Most breakpoints occurred at or very near (within a few hundred nucleotide pairs) the sites of preexisting P elements, but these elements were often lost during the rearrangement event. There were also a few cases of apparent gain of P elements. In cases in which both breakpoints of an inversion retained P elements, that inversion was capable of reverting at high frequencies to the original sequence or something close to it. This reversion occurred with sufficient precision to restore the function of a gene, held-up-b, which had been mutated by the breakpoint. However, some of the reversions had acquired irregularities at the former breakpoints that were detectable either by standard cytology or by molecular methods. The revertants themselves retained the ability to undergo further rearrangements depending on the presence of P elements. We interpret these results to rule out the simplest hypotheses of rearrangement formation that involve cointegrate structures or homologous recombination. The data provide a general picture of the rearrangement process and its possible relationship to transposition.

Choline acetyltransferase-deficient mutants of the nematode Caenorhabditis elegans.

Abstract: We have identified five independent allelic mutations, defining the gene cha-1, that result in decreased choline acetyltransferase (ChAT) activity in Caenorhabditis elegans. Four of the mutant alleles, when homozygous, lead to ChAT reductions of>98%, as well as recessive phenotypes of uncoordinated behavior, small size, slow growth and resistance to cholinesterase inhibitors. Animals homozygous for the fifth allele retain approximately 10% of the wild-type enzyme level; purified enzyme from this mutant has altered Km values for both choline and acetyl-CoA and is more thermolabile than the wild-type enzyme. These qualitative alterations, together with gene dosage data, argue that cha-1 is the structural gene for ChAT. cha-1 has been mapped to the left arm of linkage group IV and is within 0.02 map unit of the gene unc-17, mutant alleles of which lead to all of the phenotypes of cha-1 mutants except for the ChAT deficiency. Extensive complementation studies of cha-1 and unc-17 alleles reveal a complex complementation pattern, suggesting that both loci may be part of a single complex gene.

Caenorhabditis elegans deficiency mapping

Abstract: Six schemes were used to identify 80 independent recessive lethal deficiencies of linkage group (LG) II following X-ray treatment of the nematode Caenorhabditis elegans. Complementation tests between the deficiencies and ethyl methanesulfonate-induced recessive visible, lethal and sterile mutations and between different deficiencies were used to characterize the extents of the deficiencies. Deficiency endpoints thus helped to order 36 sites within a region representing about half of the loci on LG II and extending over about 5 map units. New mutations occurring in this region can be assigned to particular segments of the map by complementation tests against a small number of deficiencies; this facilitates the assignment of single-site mutations to particular genes, as we illustrate. Five sperm-defective and five oocyte-defective LG II sterile mutants were identified and mapped. Certain deficiency-by-deficiency complementation tests allowed us to suggest that the phenotypes of null mutations at two loci represented by visible alleles are wild type and that null mutations at a third locus confer a visible phenotype. A segment of LG II that is about 12 map units long and largely devoid of identified loci seems to be greatly favored for crossing over.

The cohesive population genetics of molecular drive

Abstract: The long-term population genetics of multigene families is influenced by several biased and unbiased mechanisms of nonreciprocal exchanges (gene conversion, unequal exchanges, transposition) between member genes, often distributed on several chromosomes. These mechanisms cause fluctuations in the copy number of variant genes in an individual and lead to a gradual replacement of an original family of n genes (A) in N number of individuals by a variant gene (a). The process for spreading a variant gene through a family and through a population is called molecular drive. Consideration of the known slow rates of nonreciprocal exchanges predicts that the population variance in the copy number of gene a per individual is small at any given generation during molecular drive. Genotypes at a given generation are expected only to range over a small section of all possible genotypes from one extreme (n number of A) to the other (n number of a). A theory is developed for estimating the size of the population variance by using the concept of identity coefficients. In particular, the variance in the course of spreading of a single mutant gene of a multigene family was investigated in detail, and the theory of identity coefficients at the state of steady decay of genetic variability proved to be useful. Monte Carlo simulations and numerical analysis based on realistic rates of exchange in families of known size reveal the correctness of the theoretical prediction and also assess the effect of bias in turnover. The population dynamics of molecular drive in gradually increasing the mean copy number of a variant gene without the generation of a large variance (population cohesion) is of significance regarding potential interactions between natural selection and molecular drive.

Multiple-Locus Heterozygosity and the Physiological Energetics of Growth in the Coot Clam Mulinia-Lateralis From a Natural Population

Abstract: The relationship between individual energy budgets and multiple-locus heterozygosity at six polymorphic enzyme loci was examined in Mulinia lateralis. Energy budgets were determined by measuring growth rates, rates of oxygen consumption, ammonia excretion and clearance rates. Enzyme genotypes were determined using starch gel electrophoresis. Growth rate and net growth efficiency (the ratio of energy available for growth to total energy absorbed) increased with individual heterozygosity. The positive relationship between observed growth and multiple-locus heterozygosity was associated with a negative relationship between routine metabolic costs and increasing heterozygosity. Reduction in routine metabolic costs explained 60% of the observed increased growth of more heterozygous individuals. When routine metabolic costs were standardized for differences in feeding rates, these standard metabolic costs explained 97% of the differences in growth rate. Lower standard metabolic costs, associated with increasing heterozygosity, have been proposed as a physiological mechanism for the relationship between multiple-locus heterozygosity and growth rate that has been reported for a variety of organisms, ranging in diversity from aspens to humans. This study demonstrates that reduction of standard metabolic costs, at least in clams, accounts for virtually all of the differences in growth rate among individuals of differing heterozygosity.

Meiotic and mitotic behavior of dicentric chromosomes in saccharomyces cerevisiae

Abstract: Meiotic recombination between a circular and a linear chromosome in Saccharomyces cerevisiae has been investigated. The circle was a haploid-viable derivative of chromosome III constructed by joining regions near the two chromosome ends via a recombinant DNA construction: (HMR/MAT-URA3-pBR322-MAT/HML) and was also deleted for MAL2 (which therefore uniquely marks a linear chromosome III). Recombination along chromosome III was measured for eight intervals spanning the entire length of the circular derivative. Only 25% of all tetrads from a ring/rod diploid contained four viable spores. These proved to be cases in which there was either no recombination along chromosome III or in which there were two-strand double crossovers or higher order crossovers that would not produce a dicentric chromosome.--At least half of the tetrads with three viable spores included one Ura+ Mal+ spore that was genetically highly unstable. The Ura+ Mal+ spore colonies gave rise to as many as seven genetically distinct, stable ("healed") derivatives, some of which had lost either URA3 or MAL2. Analysis of markers on chromosome III suggests that dicentric chromosomes frequently do not break during meiosis but are inherited intact into a haploid spore. In mitosis, however, the dicentric chromosome is frequently broken, giving rise to a variety of genetically distinct derivatives. We have also shown that dicentric ring chromosomes exhibit similar behavior: at least half the time they are not broken during meiosis but are broken and healed during mitosis.--The ring/rod diploid can also be used to determine the frequency of sister chromatid exchange (SCE) along an entire yeast ring chromosome. We estimate that an unequal number of SCE events occurs in approximately 15% of all cells undergoing meiosis. In contrast, the mitotic instability (and presumably SCE events) of a ring chromosome is low, occurring at a rate of about 1.2 X 10(-3) per cell division.

Genotypic Correlation and Regression in Social Groups: Multiple Alleles, Multiple Loci and Subdivided Populations

Abstract: Genotypic correlations and regressions can be estimated from multiallelic data sets either by weighting the allelic effects additively or by specifically weighting the genotypic interactions. Both methods can be extended to multiple loci, but they do not fully take into account the joint segregation patterns at the loci. These genotypic statistics have a great importance in sociobiological contexts, as they can be used for genetic descriptions of social structures. In this paper I examine the two estimation methods and show how the genotypic correlation and regression coefficients from genotypic interactions are connected to other statistics of standard population genetics; special emphasis is given to the sample-size correction when intracolony correlations from small samples were estimated. I also show how genotypic correlation and regression can be estimated in subdivided populations, both in continuous populations with isolation by distance and in populations divided into separate subpopulations. The latter analysis is an example of a more general hierarchic correlation analysis.

Mutations and Chromosomal Rearrangements Affecting the Expression of Snail, a Gene Involved in Embryonic Patterning in DROSOPHILA MELANOGASTER.

Abstract: Mutants at the snail locus are zygotically acting embryonic lethals that affect dorsoventral patterning. A comparison of seven mutant alleles shows considerable variation in expressivity and a graded effect along the dorsoventral axis: more extreme alleles result in the abnormal development of the dorsally derived ectoderm as well as the ventrally derived mesoderm, whereas weaker alleles affect only development of the mesoderm. Animals transheterozygous for different mutant alleles occasionally survive to adulthood; they frequently have missing halteres and more rarely are hemithorax. The mutant phenotype of snail is shown here to be enhanced zygotically by haploidy of two nearby regions on the second chromosome: the elbow to no-ocelli region and the interval defined by l(2)br36 and l(2)br37. It is concluded that the products of all of these genes function together in the process of specification of pattern in the embryo.

Conditioning Mutations in DROSOPHILA MELANOGASTER Affect an Experience-Dependent Behavioral Modification in Courting Males.

Abstract: One aspect of courtship in male Drosophila melanogaster has been reported to be experience dependent. Males that have courted fertilized females are virtually unresponsive to virgin females for 2-3 hr. Here, this response was utilized as an assay for the effects of conditioning mutations on experience-dependent courtship. Seven strains expressing conditioning mutations (previously isolated and characterized for learning or memory defects in an electrical shock-odor association paradigm, independent of courtship) were all found to be mutant in expression of this experience-dependent change in courtship behavior. By comparison, three control strains that were unselected for conditioning defects all expressed normal experience-dependent courtship. Other results indicate that males of the conditioning-defective strains are able to elicit necessary cues from fertilized females, yet do not then modify their courtship with virgin females. Thus, it is suggested that experience-dependent modification of courtship and the previously reported associative olfactory conditioning with electric shock share common elements of processing. The possibility that experience-dependent courtship represents adaptive behavior is discussed.

Cytogenetic analysis of a segment of the Y chromosome of Drosophila melanogaster.

Abstract: Males carrying a large deficiency in the long arm of the Y chromosome known to delete the fertility gene kl-2 are sterile and exhibit a complex phenotype: (1) First metaphase chromosomes are irregular in outline and appear sticky; (2) spermatids contain micronuclei; (3) the nebenkerns of the spermatids are nonuniform in size; (4) a high molecular weight protein ordinarily present in sperm is absent; and (5) crystals appear in the nucleus and cytoplasm of spermatocytes and spermatids. In such males that carry Ste+ on their X chromosome the crystals appear long and needle shaped; in Ste males the needles are much shorter and assemble into star-shaped aggregates. The large deficiency may be subdivided into two shorter component deficiencies. The more distal is male sterile and lacks the high molecular weight polypeptide; the more proximal is responsible for the remainder of the phenotype. Ste males carrying the more proximal component deficiency are sterile, but Ste + males are fertile. Genetic studies of chromosome segregation in such males reveal that (1) both the sex chromosomes and the large autosomes undergo nondisjunction, (2) the fourth chromosomes disjoin regularly, (3) sex chromosome nondisjunction is more frequent in cells in which the second or third chromosomes nondisjoin than in cells in which autosomal disjunction is regular, (4) in doubly exceptional cells, the sex chromosomes tend to segregate to the opposite pole from the autosomes and (5) there is meiotic drive; i.e., reciprocal meiotic products are not recovered with equal frequencies, complements with fewer chromosomes being recovered more frequently than those with more chromosomes. The proximal component deficiency can itself be further subdivided into two smaller component deficiencies, both of which have nearly normal spermatogenic phenotypes as observed in the light microscope. Meiosis in Ste + males carrying either of these small Y deficiencies is normal; Ste males, however, exhibit low levels of sex chromosome nondisjunction with either deficient Y. The meiotic phenotype is apparently sensitive to the amount of Y chromosome missing and to the Ste constitution of the X chromosome.

Healing of broken linear dicentric chromosomes in yeast

Abstract: In yeast, meiotic recombination between a linear chromosome III and a haploid-viable circular chromosome will yield a dicentric, tandemly duplicated chromosome. Spores containing apparently intact dicentric chromosomes were recovered from tetrads with three viable spores. The spore containing the dicentric inherited URA3 (part of the recombinant DNA used to join regions near the ends of the chromosome into a circle) as well as HML, HMR and MAL2 (located near the two ends of a linear but deleted from the circle). The Ura+ Mal+ colonies were highly variegated, giving rise to as many as seven distinctly different stable ("healed") derivatives, some of which were Ura+ Mal +, others Ura+ Mal- and others Ura - Mal+. The colonies were also sectored for five markers (HIS4, LEU2, CRY1, MAT and THR4) initially heterozygous in the tandemly duplicated dicentric chromosome.—Southern blot and genetic analyses have demonstrated that these stable derivatives arose from mitotic break-age of the dicentric chromosome, followed by one of several different healing events. The majority of the stable derivatives contained circular or linear chromosomes apparently resulting from homologous recombination between a broken chromosome end and a homologous region on the other end of the original dicentric duplicated chromosome. A smaller proportion of events resulted in apparently uniquely healed linear chromosomes in which the broken chromosome acquired a new telomere. In two instances we recovered chromosome III partially duplicated with a novel right end. We have also found one derivative that had also experienced rearrangement of repeated DNA sequences found adjacent to yeast telomeres.

DISTRIBUTION OF NONSTRUCTURAL VARIATION BETWEEN WHEAT CULTIVARS ALONG CHROMOSOME ARM 6Bp: EVIDENCE FROM THE LINKAGE MAP AND PHYSICAL MAP OF THE ARM

Abstract: Metaphase I (MI) pairing of homologous chromosomes in wheat intercultivar hybrids (heterohomologous chromosomes) is usually reduced relative to that within the inbred parental cultivars (euhomologous chromosomes). It was proposed elsewhere that this phenomenon is caused by polymorphism in nucleotide sequences (nonstructural chromosome variation) among wheat cultivars. The distribution of this polymorphism along chromosome arm 6Bp (=6BS) of cultivars Chinese Spring and Cheyenne was investigated. A population of potentially recombinant chromosomes derived from crossing over between telosome 6Bp of Chinese Spring and Cheyenne chromosome 6B was developed in the isogenic background of Chinese Spring. The approximate length of the Chinese Spring segment present in each of these chromosomes was assessed by determining for each chromosome the interval in which crossing over occurred (utilizing the rRNA gene region, a distal C-band and the gliadin gene region as markers). The MI pairing frequencies of these chromosomes (only the complete chromosomes were used) with the normal Chinese Spring telosome 6Bp were determined. These were directly proportional to the length of the euhomologous segment. The longer the incorporated euhomologous segment the better was the MI pairing. This provided evidence that the heterohomologous chromosomes are differentiated from each other in numerous sites distributed throughout the arm.—The comparison of the physical map of arm 6Bp with the linkage map showed a remarkable distortion of the linkage map; no crossing over was detected in the proximal 68% of the arm. A population of 49 recombinant chromosomes was assayed for recombination within the rRNA gene region, but none was detected. No new length variants of the nontranscribed spacer separating the 18S and 26S rRNA genes were detected either.