Showing papers in "Chromosoma in 1985"
TL;DR: Examination of “preimmune” serum samples from a patient who progressively developed the symptoms of scleroderma CREST over a period of several years shows that these patients make antibody species recognizing at least three distinct epitopes on C ENP-B and two on CENP-C.
Abstract: We have examined "preimmune" serum samples from a patient who progressively developed the symptoms of scleroderma CREST over a period of several years. During this period, anti-centromere antibodies (recognized by indirect immunofluorescence) appeared in the serum. Concomitant with the appearance of the anti-centromere antibodies, antibody species recognizing three chromosomal antigens in immunoblots of SDS polyacrylamide gels appeared in the patient's serum. These antigens migrate with electrophoretic mobilities corresponding to Mr = 17, 80, and 140 kilodaltons (kd). Affinity-eluted antibody fractions recognizing the antigens have been prepared from sera of three other patients. Indirect immunofluorescence labeling of mitotic cells using these antibody fractions demonstrates that the antigens are centromere components. We designate them CENP (CENtromere Protein) - A (17kd), CENP-B (80kd), and CENP-C (140kd). The three CENP antigens share antigenic determinants. Immunoblotting experiments show that these patients make antibody species recognizing at least three distinct epitopes on CENP-B and two on CENP-C. Sera from different patients contain different mixtures of the antibody species.
795 citations
TL;DR: In situ hybridization with a 125I-rDNA clone from Vicia faba was performed against Allium cepa and three strains of top onion, which represent hybrids between A. cEPa and A. fistulosum, and in principle supports the inference drawn from the Ag-NOR patterns that NORs can jump between terminal heterochromatin blocks of different Allium chromosomes in the parental species.
Abstract: In situ hybridization with a 125I-rDNA clone from Vicia faba was performed against Allium cepa and three strains of top onion, which represent hybrids between A. cepa and A. fistulosum. In principle, the labelling patterns correspond to the patterns of the silver-stained nucleolus organizing regions (NORs) in the same species. This strongly supports the inference drawn from the Ag-NOR patterns that NORs can jump between terminal heterochromatin blocks of different Allium chromosomes in the parental species A. cepa as well as in their interspecific hybrids.
317 citations
TL;DR: An aqueous spreading procedure that permits simultaneous analysis of human chromosomes by Q-banding and indirect immunofluorescence and anticentromere antibodies from an autoimmune patient is developed and it is shown that a family of structurally related human centromere proteins is detectable only at the active centromeres.
Abstract: We developed an aqueous spreading procedure that permits simultaneous analysis of human chromosomes by Q-banding and indirect immunofluorescence. Using this methodology and anticentromere antibodies from an autoimmune patient we compared the active and inactive centromeres of an isodicentric X chromosome. We show that a family of structurally related human centromere proteins (CENP-A, CENP-B, and CENP-C) is detectable only at the active centromere. These antigens therefore may be regarded both as morphological and functional markers for active centromeres.
234 citations
TL;DR: In this article, the authors examined the frequency of new chromosomal rearrangements, that appeared only in the progeny of hybrid individuals, and found that 30 times higher mutation frequency was found in hybrid males than in hybrid females.
Abstract: Introgression of a chromosome segment from Drosophila serido into the genome of its sibling D. buzzatii brought about the release of mutator potential in the hybrids. Mutator activity was determined by examining the frequency of new chromosomal rearrangements, that appeared only in the progeny of hybrid individuals. Mutation frequency was 30 times greater in the progeny of hybrid males than in that of hybrid females. There was a remarkable influence of the D. buzzatii genetic background on the frequency of production of these new rearrangements. The appearance of a new rearrangement did not depend on the genotype of the larva that bore it, but only on that of its hybrid progenitor. Among the new rearrangements there were inversions, translocations, and duplications. The number of translocations was significantly lower than that of inversions or duplications; this last type was the most frequently recorded. The distribution of the aberrations among the four major autosomes seemed to be homogeneous, although the total number of breakpoints was significantly greater in chromosome 4 than in the others. No rearrangement was found on the X chromosome. Breakpoints within three of the four affected autosomes were not randomly distributed.
180 citations
TL;DR: The organisation of the family of cross-hybridising sequences, detected by the clone p82H, is described both in the human genome and on certain chromosomes, and its relationship to known sequence families is discussed.
Abstract: Clone p82H is a human DNA sequence which hybridises in situ exclusively to the centromeric regions of all human chromosomes. It is composed of approximately 14 tandemly repeated variants of a basic 172 bp sequence, and is related to the alphoid family. The organisation of the family of cross-hybridising sequences, detected by the clone p82H, is described both in the human genome and on certain chromosomes, and its relationship to known sequence families is discussed.
161 citations
TL;DR: The somatic and meiotic chromosomes of the South American leptodactylid toads Odontophrynus americanus, Ceratophrys ornata, and C. cranwelli were analysed both with conventional staining and differential banding techniques to interpret structural heterogeneities within the quartets and octets as a “diploidization” of the polyploid karyotypes.
Abstract: The somatic and meiotic chromosomes of the South American leptodactylid toads Odontophrynus americanus, Ceratophyrys ornata, and C. cranwelli were analysed both with conventional staining and differential banding techniques. The karyotypes of O. americanus were tetraploid; those of C. ornata octaploid. Ceratophrys cranwelli is a diploid species whose karyotype displays great similarities with that of C. ornata. The high frequency of multivalent pairing configurations in the meioses of O. americanus and C. ornata indicate that these animals were of autopolyploid origin. The conventionally stained somatic chromosomes of O. americanus can be arranged into sets of four similar chromosomes (quartets); those of C. ornata, into sets of eight similar chromosomes (octets). The banding patterns revealed heterogeneity within some quartets of O. americanus, dividing each of them into two pairs of homologous chromosomes. In analogy, some octets of C. ornata can be subdivided into two quartets of chromosomes with homologous bands. These structural heterogeneities within the quartets and octets are interpreted as a "diploidization" of the polyploid karyotypes. Diploidization leads to genomes that are polyploid with respect to the amount of genetic material and diploid with respect to chromosomal characteristics and the level of gene expression. In tetraploid O. americanus, the number of nucleolus organizer regions (NORs) and their DNA content is proportional to the degree of ploidy. In contrast, up to eight NORs have been deleted in the octoploid C. ornata. These NOR losses are discussed as a possible reason for the reduction of genetic activity in polyploid genomes.
155 citations
TL;DR: Results demonstrate that the nucleolus is formed by fusion of prenucleolar structures whose biochemical composition resembles the matureucleolus, since PNBs contain at least two of the major nucleolar proteins.
Abstract: A time course study was conducted on nucleologenesis after release from a mitotic block in the presence and absence of actinomycin D to determine the composition and fate of prenucleolar bodies (PNBs). Prenucleolar bodies, whether naturally occurring or induced by actinomycin D treatment, stain with silver and contain phosphoproteins B23 and C23, two of the major proteins of the interphase nucleolus as determined by double label immunofluorescence with specific antibodies. The nucleolus is formed by fusion of PNBs, which subsequently “reorganize” and form internal fibrillar and peripheral granular regions. Actinomycin D prevents fusion of PNBs, which are then randomly dispersed throughout the nucleus but they still contain proteins B23 and C23. These results demonstrate that the nucleolus is formed by fusion of prenucleolar structures whose biochemical composition resembles the mature nucleolus, since PNBs contain at least two of the major nucleolar proteins.
125 citations
TL;DR: The pathways of endoreplication in three cell types, salivary gland, nurse, and follicle cells, share basic features of DNA replication, and differ primarily in the extent of association of the duplicated chromatids.
Abstract: In the nurse cells of Drosophila, nuclear DNA is replicated many times without nuclear division. Nurse cells differ from salivary gland cells, another type of endoreplicated Drosophila cell, in that banded polytene chromosomes are not seen in large nurse cells. Cytophotometry of Feulgen stained nurse cell nuclei that have also been labeled with 3H-thymidine shows that the DNA contents between S-phases are not doublings of the diploid value. In situ hybridization of cloned probes for 28S+18S ribosomal RNA, 5S RNA, and histone genes, and for satellite, copia, and telomere sequences shows that satellite and histone sequences replicate only partially during nurse cell growth, while 5S sequences fully replicate. However, during the last nurse cell endoreplication cycle, all sequences including the previously under-replicated satellite sequences replicate fully. In situ hybridization experiments also demonstrate that the loci for the multiple copies of histone and 5S RNA genes are clustered into a small number of sites. In contrast, 28S+18S rRNA genes are dispersed. We discuss the implications of the observed distribution of sequences within nurse cell nuclei for interphase nuclear organization. — In the ovarian follicle cells, which undergo only two or three endoreplication cycles, satellite, histone and ribosomal DNA sequences are also found by in situ hybridization to be underrepresented; satellite sequences may not replicate beyond their level in 2C cells. Hence the pathways of endoreplication in three cell types, salivary gland, nurse, and follicle cells, share basic features of DNA replication, and differ primarily in the extent of association of the duplicated chromatids.
106 citations
TL;DR: Analysis of D15Z1 indicated that this sequence is composed of tandemly arranged imperfect repeats of the consensus 5′ AATGG 3′ similar to previously identified satellite III sequences and that it makes up the major fraction of the heterochromatin of this chromosome.
Abstract: We have isolated a repetitive 1.8 kb Kpnl DNA sequence which is amplified in the homogeneously staining regions of a human melanoma cell line. Under low stringency conditions this sequence (D15Z1) hybridized in situ to the centromeric heterochromatin of chromosomes 1, 9, 15p, 16, and distal Yq as well as to the the short arms of the other acrocentric chromosomes. Under conditions of high stringency, labelling was predominantly on the short arm of chromosome 15. D15Z1 was shown to be present at approximately 3,000 copies per haploid genome and organized in long tandem arrays showing restriction site heterogeneity. Sequences homologous to D15Z1 were highly enriched in the less dense shoulder region of a Ag+—Cs2SO4 gradient. Analysis of D15Z1 indicated that this sequence is composed of tandemly arranged imperfect repeats of the consensus 5′ AATGG 3′ similar to previously identified satellite III sequences. Digestion of D15Z1 with HinfI resulted in a series of restriction fragments making up a subset of the HinfI ladder components of satellites III and IV. These data suggest that D15Z1 represents a chromosome 15 specific domain of human satellites III or IV and that it makes up the major fraction of the heterochromatin of this chromosome. Possible relationships between this sequence and the cytochemical staining properties of human chromosomes with distamycin A/DAPI, D280/170, and antiserum to 5-methylcytosine are discussed.
104 citations
TL;DR: It is concluded that chromosome movement can cause reorientation to begin and that in the changes which follow, an unstable attachment of kinetochore microtubules to the spindle plays a major role.
Abstract: The relationship between chromosome movement and microtubules was explored by combining micromanipulation of living grasshopper spermatocytes with electron microscopy. We detached chromosomes from the spindle and placed them far out in the cytoplasm. Soon, the chromosomes began to move back toward the spindle and the cells were fixed at a chosen moment. The microtubules seen in three-dimensional reconstructions were correlated with the chromosome movement just prior to fixation. Before movement began, detached chromosomes had no kinetochore microtubules or a single one at most. Renewed movement was always accompanied by the reappearance of kinetochore microtubules; a single kinetochore microtubule appeared to suffice. Chromosome movements and kinetochore microtubule arrangements were unusual after reattachment, but their relationship was not: poleward forces, parallel to the kinetochore microtubule axis (as in normal anaphase), would explain the movement, however odd. The initial arrangement of kinetochore microtubules would have led to aberrant chromosome distribution if it persisted, but instead, reorientation to the appropriate arrangement always followed. Observations on living cells permitted us to place in sequence the kinetochore microtubule arrangements seen in fixed cells, revealing the microtubule transformations during reorientation. From the sequence of events we conclude that chromosome movement can cause reorientation to begin and that in the changes which follow, an unstable attachment of kinetochore microtubules to the spindle plays a major role.
100 citations
TL;DR: It was confirmed that the pattern of chromosome banding induced by AluI, MboI, HaeIII, DdeI, RsaI, and HinfI is characteristic for each endonuclease, and several restriction banding polymorphisms that were not found by conventional C-banding were detected, indicating that there may be a range of variability in the frequency and distribution of restriction sites in homologous chromosome regions.
Abstract: The mechanism of chromosome banding induced by restriction endonucleases was analyzed by measuring the amount of radioactivity extracted from [14C]thymidine-labeled chromosomes digested first with restriction enzymes and subsequently with proteinase K and DNase I. Restriction enzymes with a high frequency of recognition sites in the DNA produced a large number of short DNA fragments, which were extracted from chromosomes during incubation with the enzyme. This loss of DNA resulted in decreased chromosomal staining, which did not occur in regions resistant to restriction enzyme digestion and thus led to banding. Subsequent digestion of chromosomes with proteinase K produced a further loss of DNA, which probably corresponded to long fragments retained in the chromosome by the proteins of fixed chromatin. Restriction enzymes induce chromatin digestion and banding in G1 and metaphase chromosomes, and they induce digestion and the appearance of chromocenters in interphase nuclei. This suggests that the spatial organization and folding of the chromatin fibril plays little or no role in the mechanism of chromosome banding. It was confirmed that the pattern of chromosome banding induced by AluI, MboI, HaeIII, DdeI, RsaI, and HinfI is characteristic for each endonuclease. Moreover, several restriction banding polymorphisms that were not found by conventional C-banding were detected, indicating that there may be a range of variability in the frequency and distribution of restriction sites in homologous chromosome regions.
TL;DR: This work investigates the organisation, nucleotide sequence, and chromosomal distribution of a tandemly repeated, satellite DNA from Allium cepa (Liliaceae), and presents the nucleotide sequences of three repeats, which differ at a large number of positions.
Abstract: We have investigated the organisation, nucleotide sequence, and chromosomal distribution of a tandemly repeated, satellite DNA from Allium cepa (Liliaceae). The satellite, which constitutes about 4% of the A. cepa genome, may be resolved from main-band DNA in antibiotic-CsCl density gradients, and has a repeat length of about 375 base pairs (bp). A cloned member of the repeat family hybridises exclusively to chromosome telomeres and has a non-random distribution in interphase nuclei. We present the nucleotide sequences of three repeats, which differ at a large number of positions. In addition to arrays made up of 375-bp repeats, homologous sequences are found in units with a greater repeat length. This divergence between repeats reflects the heterogeneity of the satellite determined using other criteria. Possible constraints on the interchromosomal exchange of repeated sequences are discussed.
TL;DR: Comparative fluorescence studies on the chromosome of ten species of acridid grasshoppers indicate that the only regions to fluoresce differentially are those that C-band, and that structural rearrangements that lead to the relocation of centric C-bands, either within or between species, may also be accompanied by a change in fluorescence behaviour.
Abstract: Comparative fluorescence studies on the chromosome of ten species of acridid grasshoppers, with varying amounts and locations of C-band positive heterochromatin, indicate that the only regions to fluoresce differentially are those that C-band Within a given species there is a marked tendency for groups of chromosomes to accumulate heterochromatin with similar fluorescence behaviour at similar sites This applies to all three major categories of heterochromatin — centric, interstitial and telomeric Different sites within the same complement, however, tend to have different fluorescence properties In particular, centric C-bands within a given species are regularly distinguishable in their behaviour from telomeric C-bands Different species, on the other hand, may show distinct forms of differential fluorescence at equilocal sites These varying patterns of heterochromatin heterogeneity, both within and between species, indicate that whatever determines the differential response to fluorochromes has tended to operate both on an equilocal basis and in a concerted fashion This is reinforced by the fact that structural rearrangements that lead to the relocation of centric C-bands, either within or between species, may also be accompanied by a change in fluorescence behaviour
TL;DR: It is shown that 5-aza-C causes a dramatic increase in the DNase-I sensitivity of the entire inactive X chromosome of female G. gerbillus cells and this increase in nuclease sensitivity correlates with a large shift in the time of replication of the inactive X chromosomes from late S phase to early S phase.
Abstract: We have examined the effect of 5-azacytidine (5-aza-C) induced hypomethylation of DNA on the time of replication and DNase I sensitivity of the X chromosomes of female Gerbillus gerbillus (rodent) lung fibroblast cells. Using in situ nick translation to visualise the potential state of activity of large regions of metaphase chromosomes we show that 5-aza-C causes a dramatic increase in the DNase-I sensitivity of the entire inactive X chromosome of female G. gerbillus cells and this increase in nuclease sensitivity correlates with a large shift in the time of replication of the inactive X chromosome from late S phase to early S phase. These effects of 5-aza-C on the inactive X chromosome are associated with a 15% decrease in DNA methylation. Our results indicate that DNA methylation concomitantly affects both the time of replication and the chromatin conformation of the inactive X chromosome.
TL;DR: In this article, the relative amount of DNA in defined segments of salivary gland chromosomes of Drosophila melangogaster from the Oregon R stock was determined by autoradiography.
Abstract: The relative amount of DNA in defined segments of salivary gland chromosomes of Drosophila melangogaster from the Oregon R stock was determined by autoradiography. The data obtained were then used to estimate the possible correlation between DNA content and the degree of manifestation of charcters such as weak-point behavior, late replication, strong synapsis, breaks of chromosome rearrangements, hybridization with cRNA, and localization of mobile elements. Of 380 regions investigated 274 have showed deviations in the degree of manifestation of these features from that predicted on the basis of the DNA content of these regions. Regions, previously shown to consist of intercalary heterochromatin (IH, Zhimulev et al. 1982), were found to have a significantly higher frequency of the simultaneous manifestation of several of the above-mentioned features, with the exception of localization of mobile elements. These findings support the earlier suggestion that a high frequency and a simultaneous manifestation of IH features depend on some peculiarities of the molecular organization of IH regions, but not on a high DNA content.
TL;DR: Zygotene synaptonemal complex formation in rye is characterised by existence of a bouquet, with telomeric pairing initiation earliest, and new pairing initiations occur almost until the end of zygotenes.
Abstract: A spreading technique was used to allow ultrastructural analysis of seventeen zygotene nuclei of rye (Secale cereale) Twenty pachytene nuclei were also examined Lateral element lengths of the haploid complements decreased from 742 μm at the beginning of zygotene to 451 μm at the end of zygotene Variation in pachytene synaptonemal complex lengths was also noted Zygotene synaptonemal complex formation in rye is characterised by: (1) existence of a bouquet, with telomeric pairing initiation earliest; (2) multiple sites of initiation in each bivalent (maximum of 76 synaptonemal complex segments seen in one nucleus); (3) the potential number of pairing initiation sites may be higher (the average spacing of 442 μm would allow approximately 160 sites per nucleus); (4) new pairing initiations occur almost until the end of zygotene; (5) initiation of new synaptonemal complexes and extension of existing synaptonemal complexes occur simultaneously A simple zipping up of a few initiation sites is not the case in rye Pairing in different bivalents of a nucleus is not completely synchronised, and the NOR in particular is often late to pair Interlocking of lateral elements and synaptonemal complexes may lead to delayed completion of pairing in portions of bivalents, but interlocks are ultimately resolved This resolution may involve breakage and rejoining of lateral elements
TL;DR: Heterochromatin in these taxa is thus highly differentiated, exhibiting heterogeneity in staining characteristics, and presumably in underlying DNA sequences, both across the genome within a given chromosomal complement as well as among the different karyotypic races and species of the bottae group of pocket gophers.
Abstract: Heterochromatin is a dominant component of the genome in the bottae group of the pocket gopher genus Thomomys, having had a major role in the karyotypic evolution of member species. Heterochromatin characteristics of two subspecies of T. bottae and one of T. umbrinus were examined with fluorochrome dyes identifying presumptive GC- and AT-rich regions. In two karyotypic forms of T. b. fulvus and in T. umbrinus, chromatin that fluoresces brightly with chromomycin A3 is also C-band positive, although not all heterochromatin fluoresces. However, in T. b. bottae, only euchromatic regions fluoresce brightly with chromomycin. Fluorescence patterns produced with DAPI are the reverse of the chromomycin banding in all karyotypic forms. Heterochromatin in these taxa is thus highly differentiated, exhibiting heterogeneity in staining characteristics, and presumably in underlying DNA sequences, both across the genome within a given chromosomal complement as well as among the different karyotypic races and species of the bottae group of pocket gophers.
TL;DR: The occurrence of peak levels of m-rec and recombination activity in extracts from cells in early pachytene points strongly to that stage as the time at which the enzymatic phase of recombination occurs.
Abstract: RecA-like proteins have been purified from somatic and meiotic cells of mouse and lily. The rec proteins have been designated "s-rec" and "m-rec" to indicate their respective tissues of origin. The two proteins differ in molecular weight and in their response to temperature, the latter being consistent with the optimal temperature for physiological function of their tissues of origin. There is a major increase in m-rec protein with the entry of cells into meiosis, the peak of activity being early pachytene. Extracts of the cells and also those of yeast (Saccharomyces cerevisiae) have been prepared that have the capacity to catalyze homologous recombination. These extracts behave similarly to the m-rec proteins upon entry of cells into meiosis. Yeast transferred to sporulation medium displays a 100-fold increase in the recombination activity of the extract at about the time of entry into meiosis. The occurrence of peak levels of m-rec and recombination activity in extracts from cells in early pachytene points strongly to that stage as the time at which the enzymatic phase of recombination occurs.
TL;DR: It is concluded that DNA replication in polytene cells is controlled by modifications of the mechanism operating in dividing cells, where control is sequence autonomous, and not by a “doubling of DNA” mechanism.
Abstract: In dividing cells, each sequence replicates exactly once in each S-phase, but in cells with polytene chromosomes, some sequences may replicate more than once or fail to replicate during S-phase. Because of this differential replication, the control of replication in polytene cells must have some unusual features. Dennhofer (1982a) has recently concluded that the total DNA content of the polytene cells of Drosophila salivary glands exactly doubles in each S-phase. This observation, along with previous studies demonstrating satellite underreplication in salivary gland cells, led us to consider the hypothesis that there is a "doubling of DNA" mechanism for the control of DNA replication in polytene cells. With this mechanism, a doubling of DNA content, rather than the replication of each sequence, would signal the end of a cycle of DNA replication. To test this hypothesis, we have reinvestigated the replication of several sequences (satellite, ribosomal, histone and telomere) in salivary gland cells using quantitative in situ hybridization. We find that underreplication of some sequences does occur. In addition we have repeated Dennhofer's cytophotometric and labeling studies. In contrast to Dennhofer, we find that the total DNA contents of nonreplicating nuclei do reflect this partial replication, in accord with Rudkin's (1969) result. We conclude that DNA replication in polytene cells is controlled by modifications of the mechanism operating in dividing cells, where control is sequence autonomous, and not by a "doubling of DNA" mechanism. In situ hybridization to unbroken salivary gland nuclei reveals the distribution of specific sequences. As expected, satellite, histone and 5S sequences are usually in a single cluster.(ABSTRACT TRUNCATED AT 250 WORDS)
TL;DR: Describing at the nucleotide level a 4.8-kilobase pair segment of the third chromosome of Droophila melanogaster, which contains a cluster of three chorion genes, finds the highest number of dyad symmetries, remiiscent of sequences that function as viral replication origins, is found associated with the T- and A-rich regions between genes s18-1 and s15-1.
Abstract: We have characterized at the nucleotide level a 4.8-kilobase pair segment of the third chromosome of Droophila melanogaster, which contains a cluster of three chorion genes, s 18-1, s 15-1 and s 19-1. These genes are tandemly oriented and share the same basic organization: a small and a large exon separated by a short intron in the signal peptide region. In the coding region, limited similarities at the DNA and protein level suggest a common but distant evolutionary origin. The flanking sequences were searched for elements that might be involved in controlling the tissue-specific and temporally regulated expression and the selective amplification of the chorion genes. A good candidate for a cis-regulatory element is the hexamer, TCACGT, which is found in all three genes in a highly significant position, 23 to 27 nucleotides upstream of the TATA-box, accompanied by additional, less exact similarities. Palindromes and short inverted repeats that are found in the vicinity of their complement are non-uniformly distributed: they are most concentrated in the 3′ flanking part of all three genes, in and near regions of unusually high A and T content. The highest number of dyad symmetries, remiiscent of sequences that function as viral replication origins, is found associated with the T- and A-rich regions between genes s18-1 and s15-1.
TL;DR: It is suggested that sex-linked complex heterozygosity is also found in a number of species of social termites, and may have adaptive significance for a colonial lifestyle.
Abstract: In the vast majority of spider species studied to date, the karyotype is homogeneous in morphology and exclusively telocentric. The sex-determining system consists of one to three X chromosomes in the male and, correspondingly, two to six in the female. This is the case in species of huntsman spiders belonging to the genera Heteropoda (2n=40+3X), Isopoda, Olios, and Pediana (2n=40+3X) and some populations of the colonial species Delena cancerides (2n=40+3X). In other populations of D. cancerides, wholesale fusion of the karyotype has occurred, reducing the standard huntsman karyotype of 43 telocentric chromosomes to 21 metacentrics and 1 telocentric. Eight of the centric fusion products, including an X-autosome fusion, are maintained in the heterozygous condition in males and, with the single telocentric, form a chain of nine chromosomes at meiosis. The two complexes comprising the chain behave as neo-X and neo-Y chromosomes, and thus the ancestral X1X2X3♂∶X1X1X2X2X3X3♀ sex-determining system has been converted to a system of six X and four Y chromosomes in the male and twelve X chromosomes in the female. Since sex-linked complex heterozygosity is also found in a number of species of social termites, it is suggested that such heterozygosity may have adaptive significance for a colonial lifestyle. Breakdown products of the chain of nine are present in specimens of D. cancerides from Canberra and these appear to represent hybrid products between the 2n=22 and 2n=43 forms. Hybridisation may also have been involved in the origin of the chain-forming races.
TL;DR: Comparisons were made between two kinds of tetraploids derived from the hybrid Lolium temulentum x L. perenne, in which some homologous bivalents are not the product of resolution of multivalents but result from two-by-two pairing from the beginning of zygotene.
Abstract: The chromosomes of Lolium temulentum are longer and contain on average 50% more nuclear DNA than the chromosomes of L. perenne. In the hybrid, despite the difference in length and DNA content, pairing between the homoeologous chromosomes at pachytene is effective and the chiasma frequency at first metaphase in pollen mother cells is high, about 1.6 per bivalent, comparable to that in the L. perenne parent. Electron microscopic observations from reconstructed nuclei at pachytene show that synaptonemal complex (SC) formation in 40% of bivalents is “perfect,” complete and continuous from telomere to telomere. In others, SCs extend from telomere to telomere but incorporate lateral component loops in interstitial chromosome segments. Even in these bivalents, however, pairing is effective in the sense of chiasma formation. The capacity to form “perfect” SCs is achieved by an adjustment of chromosome length differences both before and during synapsis. “Perfect” pairing and SC formation is commoner within the larger bivalents of the complement. At zygotene, in contrast to pachytene, pairing is not confined to homoeologous chromosomes. On the contrary there is “illegitimate” pairing between non-homologous chromsomes resulting in multivalent formation. There must, therefore, be a mechanism operative between zygotene and pachytene that corrects and modifies associations in such a way as to restrict the pairing to bivalents comprised of strictly homoeologous chromosomes. Such a correction bears comparison with that known to apply in allopolyploids. In the hybrid and in the L. perenne parent also, certain specific nucleolar organisers are inactivated at meiosis.
TL;DR: Prometaphase I chromosome behavior was examined in wild-type Drosophila melanogaster primary spermatocytes and results suggest that most chromosome motions can be explained by poleward forces acting on or through kinetochore microtubules that span the distance between the kinetchore and the vicinity of a pole.
Abstract: Prometaphase I chromosome behavior was examined in wild-type Drosophila melanogaster primary spermatocytes. Cine analysis of live cells reveals that bivalents exhibit complex motions that include (1) transient bipolar orientations, (2) simultaneous reorientation of homologous kinetochores, (3) movements not parallel to the spindle axis, and (4) movement along the nuclear membrane. --Kinetochores and kinetochore microtubule have been analyzed for bivalents previously studied in life. The results suggest that most chromosome motions (complex though they may be) can be explained by poleward forces acting on or through kinetochore microtubules that span the distance between the kinetochore and the vicinity of a pole. The results also suggest that the majority of short kinetochore microtubules may be remnants of previous microtubule-mediated associations between a kinetochore and a pole.
TL;DR: It is shown that a DNA segment, consisting of four tandemly repeated ry gene copies and interspersed by material which includes P DNA, forms a complex of loose chromatin in which, however, four bands can be resolved.
Abstract: Data are presented of electron microscopic (EM) analysis of consecutive developmental stages of Drosophila melanogaster complex puffs, formed as a result of simultaneous decondensation of several bands. EM mapping principles proposed by us permitted more exact determination of the banding patterns of 19 regions in which 31 puffs develop. It is shown that 20 of them develop as a result of synchronous decondensation of two bands, 7 of three and 4 of one band. Three cases of two-band puff formation when one or both bands undergo partial decondensation are descirbed. In the 50CF, 62CE, 63F and 71CF regions puffing zones are located closely adjacent to each other but the decondensation of separate band groups occurs at different puff stages (PS). These data are interpreted as activation of independently regulated DNA sequences. The decondensation of two or three adjacent bands during formation of the majority of the puffs occurs simultaneously in the very first stages of their development. It demonstrates synchronous activation of the material of several bands presumably affected by a common inductor. Bands adjacent to puffing centres also lose their clarity as the puff develops, probably due to “passive” decondensation connected with puff growth. The morphological data obtained suggest a complex genetic organisation of many puffs.
TL;DR: The complete nucleotide sequence of this “amplification control element” and of genes encoding the chorion structural proteins s18-1 and s15-1, which are contained within it are presented and Sequences that may be involved in the regulation of chorions gene amplification and expression are identified.
Abstract: During Drosophila oogenesis, two clusters of chorion genes and their flanking DNA sequences undergo amplification in the ovarian follicle cells. Amplification results from repeated rounds of initiation and bidirectional replication within the chorion gene regions, possibly from a single origin, producing nested replication forks. Previously we have shown that following reintroduction into the Drosophila genome, a specific 3.8 kilobase pair DNA segment from the amplified third chromosome domain could induce developmentally regulated amplification at its site of insertion. Here we present the complete nucleotide sequence of this “amplification control element” and of genes encoding the chorion structural proteins s18-1 and s15-1, which are contained within it. Sequences that may be involved in the regulation of chorion gene amplification and expression are identified.
TL;DR: A model for Sce induction is proposed involving a single-strand break or gap as the initial requirement for SCE initiation at the replicating fork, resulting in enhanced SCE levels during the second cycle of BrdU incorporation.
Abstract: 5-Bromodeoxyuridine (BrdU)-induced sister chromatid exchanges (SCEs) are mainly determined during replication on a BrdU-substituted template. The BrdU, once incorporated, is rapidly excised as uracil (U), and the gap is repaired with the incorporation of BrdU from the medium, which leads to further repair. During the second S period in BrdU medium, this process continues as the strand acts as template. Experiments suggest that 3-amino-benzamide (3AB) delays the ligation of the gaps formed after U excision, resulting in enhanced SCE levels during the second cycle of BrdU incorporation. When normal templates of G1 cells are treated before BrdU introduction with methyl methanesulphonate (MMS), 3AB in the first cycle doubles the MMS-induced SCEs but has no effect on them during the second cycle. When the BrdU-substituted template is treated with MMS in G1 of the second cycle, 3AB again doubles the SCEs due to MMS and also enhances the SCEs resulting from delays in ligation of the gaps following U excision in the BrdU-substituted template. The repair processes of MMS lesions that are sensitive to 3AB and lead to SCEs take place rapidly, while the repair process of late repairing lesions that lead to SCEs appear to be insensitive to 3AB. A model for SCE induction is proposed involving a single-strand break or gap as the initial requirement for SCE initiation at the replicating fork. Subsequent events represent natural stages in the repair process of a lesion, ensuring replication without loss of genetic information. G1 cells treated with methylnitrosourea (MNU) and grown immediately in BrdU medium rapidly lose the O6-methylguanine from their DNA and the rate of loss is BrdU-dose dependent. The rapid excision of the U lesions can explain the effect of BrdU concentration on SCE reduction following both MNU or MMS treatment.
TL;DR: Electron microscopic examination of silver-stained chromosomes indicated that the lateral elements of the synaptonemal complexes were prominently stained, and terminal attachment plaques were apparent, and a translocation quadrivalent could be recognized easily in the light microscope using these methods.
Abstract: We have taken advantage of the synchronous meiotic process in the basidiomycete Coprinus cinereus to develop a simple and rapid method to selectively stain meiotic chromosomes and nucleoli in this fungus without prior removal of the cell wall. Electron microscopic examination of these silver-stained chromosomes indicated that the lateral elements of the synaptonemal complexes were prominently stained, and terminal attachment plaques were apparent. We found that a translocation quadrivalent could be recognized easily in the light microscope using these methods. The procedures appear suitable for the characterization of chromosome rearrangements in this small genome, and should facilitate cytogenetic analysis in this fungus.
TL;DR: In Chironomus thumMI thummi a Balbiani ring-like structure is induced at the right end of chromosome III by heat shock, andTelomere-specific clones obtained by microcloning this chromosome region showed some sequence divergence, which seemed to increase when different telomeres were compared.
Abstract: In Chironomus thummi thummi a Balbiani ring-like structure is induced at the right end of chromosome III by heat shock. Telomere-specific clones were obtained by microcloning this chromosome region. One of the recombinant clones, λCthS, which had an insert of 176 base pairs (bp), hybridized in situ to all the chromosome ends of C. th. thummi except one on the acrocentric chromosome IV. Homologous sequences were also present at two interstitial sites on chromosome II. Southern analysis of genomic DNA indicated that the 176-bp insert represents a basic repeating unit clustered in tandem arrays. Around 1000 copies of this sequence were located in each telomere. The repeats were homogeneous in size but showed some sequence divergence, which seemed to increase when different telomeres were compared. At the nucleotide sequence level the most noticeable feature was the presence of tracts of adenine alternating with tracts of thymine. In situ hybridzation to chromosomal RNA showed that the 176-bp repeat was complementary to the RNA transcribed at the T-BRIII region, and occasionally, to the RNA transcribed at some of the other telomeres during heat shock. Northern blot analysis of total RNA showed that a transcript larger than 10 kilobase pairs (kb) and complementary to the teloeric insert was present in heat-shocked as well as in untreated cells. However, at most only a very weak RNA hybridization was seen at the telomeres of untreated animals.
TL;DR: These autosomal repeated DNA domains, like their Y chromosome counterparts, consist of a complex mixture of repeated DNA elements interspersed among each other in ways that lead to defined periodicities.
Abstract: The human Y chromosome contains a group of repeated DNA elements, identified as 3.4-kilobase pair (kb) fragments in Hae III digests of male genomic DNA, which contain both Y-specific and non-Y-specific sequences. We have used these 3.4-kb Hae III Y fragments to explore the organizational properties and chromosomal distribution of the autosomal homologs of the non-Y-specific (NYS) 3.4-kb Hae III Y elements. Three distinct organizations, termed domains, have been identified and shown to have major concentrations on separate chromosomes. We have established that domain K is located on chromosome 15 and domain D on chromosome 16 and suggested that domain R is on chromosome 1. Our findings suggest that each domain is composed of a tandemly arrayed cluster of a regularly repeating unit containing two sets of repeated sequences: one that is homologous to the NYS 3.4-kb Hae III Y sequences and one that does not cross-react with the 3.4-kb Hae III Y repeats. Thus, these autosomal repeated DNA domains, like their Y chromosome counterparts, consist of a complex mixture of repeated DNA elements interspersed among each other in ways that lead to defined periodicities. Although each of the three identified autosomal domains cross-reacts with 3.4-kb Hae III Y fragments purified from genomic DNA, the length periodicities and sequence content of the autosomal domains are chromosome specific. The organizational properties and chromosomal distribution of these NYS 3.4-kb Hae III homologs seem inconsistent with stochastic mechanisms of sequence diffusion between chromosomes.
TL;DR: The results suggest that the amplified c-myc appeared first as DMs and was subsequently transposed to engender HSRs on an X chromosome, suggesting that the initial COLO 320 tumor cell may have acquired two “early replicating” X chromosomes and lost the “late replicating" X.
Abstract: Cell lines (COLO 320 DM and COLO 320 HSR), established from a human neuroendocrine tumor, contain an amplified cellular oncogene (c-myc). We have previously shown that the homogeneously staining regions (HSRs) of a marker chromosome in the COLO 320 HSR cells that evolved in culture from COLO 320 DM cells contain amplified c-myc. Molecular hybridization in situ has now been used to demonstrate that the HSRs are on both arms of what was once an X chromosome. We also show that amplified c-myc copies are present in the isolated double minute chromosomes (DMs) of the COLO 320 DM cells that were characteristic of the tumor cells initially established from the patient. The results suggest that the amplified c-myc appeared first as DMs and was subsequently transposed to engender HSRs on an X chromosome. The initial COLO 320 tumor cell may have acquired two "early replicating" (i.e., active) X chromosomes and lost the "late replicating" (i.e., inactive) X.