Showing papers on "Gene published in 1983"
TL;DR: In four of five patients studied, representing two histological types of cancer, substantial hypomethylation was found in genes of cancer cells compared with their normal counterparts, and was progressive in a metastasis from one of the patients.
Abstract: It has been suggested that cancer represents an alteration in DNA, heritable by progeny cells, that leads to abnormally regulated expression of normal cellular genes; DNA alterations such as mutations, rearrangements and changes in methylation have been proposed to have such a role. Because of increasing evidence that DNA methylation is important in gene expression (for review see refs 7, 9-11), several investigators have studied DNA methylation in animal tumours, transformed cells and leukaemia cells in culture. The results of these studies have varied; depending on the techniques and systems used, an increase, decrease, or no change in the degree of methylation has been reported. To our knowledge, however, primary human tumour tissues have not been used in such studies. We have now examined DNA methylation in human cancer with three considerations in mind: (1) the methylation pattern of specific genes, rather than total levels of methylation, was determined; (2) human cancers and adjacent analogous normal tissues, unconditioned by culture media, were analysed; and (3) the cancers were taken from patients who had received neither radiation nor chemotherapy. In four of five patients studied, representing two histological types of cancer, substantial hypomethylation was found in genes of cancer cells compared with their normal counterparts. This hypomethylation was progressive in a metastasis from one of the patients.
2,358 citations
TL;DR: The chromosomal localization of the Huntington's disease gene is the first step in using recombinant DNA technology to identify the primary genetic defect in this disorder.
Abstract: Family studies show that the Huntington's disease gene is linked to a polymorphic DNA marker that maps to human chromosome 4. The chromosomal localization of the Huntington's disease gene is the first step in using recombinant DNA technology to identify the primary genetic defect in this disorder.
2,211 citations
TL;DR: The provirus of ATLV integrated in DNA of leukemia T cells from a patient with ATL was molecularly cloned and the complete nucleotide sequence of the proviral genome was determined.
Abstract: Human retrovirus adult T-cell leukemia virus (ATLV) has been shown to be closely associated with human adult T-cell leukemia (ATL) [Yoshida, M., Miyoshi, I. & Hinuma, Y. (1982) Proc. Natl. Acad. Sci. USA 79, 2031-2035]. The provirus of ATLV integrated in DNA of leukemia T cells from a patient with ATL was molecularly cloned and the complete nucleotide sequence of 9,032 bases of the proviral genome was determined. The provirus DNA contains two long terminal repeats (LTRs) consisting of 755 bases, one at each end, which are flanked by a 6-base direct repeat of the cellular DNA sequence. The nucleotides in the LTR could be arranged into a unique secondary structure, which could explain transcriptional termination within the 3' LTR but not in the 5' LTR. The nucleotide sequence of the provirus contains three large open reading frames, which are capable of coding for proteins of 48,000, 99,000, and 54,000 daltons. The three open frames are in this order from the 5' end of the viral genome and the predicted 48,000-dalton polypeptide is a precursor of gag proteins, because it has an identical amino acid sequence to that of the NH2 terminus of human T-cell leukemia virus (HTLV) p24. The open frames coding for 99,000- and 54,000-dalton polypeptides are thought to be the pol and env genes, respectively. On the 3' side of these three open frames, the ATLV sequence has four smaller open frames in various phases; these frames may code for 10,000-, 11,000-, 12,000-, and 27,000-dalton polypeptides. Although one or some of these open frames could be the transforming gene of this virus, in preliminary analysis, DNA of this region has no homology with the normal human genome.
1,519 citations
TL;DR: It is shown here that a DNA domain detectable by partial homology to the myc oncogene is amplified up to 140-fold in cell lines derived from different human neuroblastomas and in a neuroblastoma tumour, but not in other tumour cells showing cytological evidence for gene amplification.
Abstract: Amplified cellular genes in mammalian cells frequently manifest themselves as double minute chromosomes (DMs) and homogeneously staining regions of chromosomes (HSRs). With few exceptions both karyotypic abnormalities appear to be confined to tumour cells. All vertebrates possess a set of cellular genes homologous to the transforming genes of RNA tumour viruses, and there is circumstantial evidence that these cellular oncogenes are involved in tumorigenesis. We have recently shown that DMs and HSRs in cells of the mouse adrenocortical tumour Y1 and an HSR in the human colon carcinoma COLO320 contain amplified copies of the cellular oncogenes c-Ki-ras and c-myc, respectively. Both DMs and HSRs are found with remarkable frequency in cells of human neuroblastomas. We show here that a DNA domain detectable by partial homology to the myc oncogene is amplified up to 140-fold in cell lines derived from different human neuroblastomas and in a neuroblastoma tumour, but not in other tumour cells showing cytological evidence for gene amplification. By in situ hybridization we found that HSRs are the chromosomal sites of the amplified DNA. The frequency with which this amplification appears in cells from neuroblastomas and its apparent specificity raise the possibility that one or more of the genes contained within the amplified domain contribute to tumorigenesis.
1,366 citations
TL;DR: The complete nucleotide sequence of bacteriophage T7 DNA, 39,936 base-pairs, has been determined and the amino acid sequences and compositions predicted for all of the T7 proteins (except the proteins produced by frameshifting) are given.
1,289 citations
TL;DR: It is proposed that this tissue-specific enhancer contributes to the activation of somatically rearranged immunoglobulin variable region genes and possibly to abnormal expression of other genes (e.g. c-myc) that become translocated to its domain of influence.
1,285 citations
TL;DR: The polyoma virus middle-T and the T24 Harvey ras1 genes are individually unable to transform primary baby rat kidney cells and separate establishment and transforming functions are required for oncogenic transformation of primary cells in culture.
Abstract: The polyoma virus middle-T and the T24 Harvey ras1 genes are individually unable to transform primary baby rat kidney cells. Adenovirus early region 1A provides functions required by these genes to transform primary cells following DNA-mediated gene transfer. These results suggest that separate establishment and transforming functions are required for oncogenic transformation of primary cells in culture.
1,245 citations
TL;DR: Analysis of the DNA sequences of the 5' end of the clones demonstrated that although alpha- and gamma-actin genes start with a methionine codon, it is likely that post-translational removal of cysteine in addition to methionines accompanies alpha-actIn synthesis but not beta- and Gamma-Actin synthesis.
Abstract: cDNA clones encoding three classes of human actins have been isolated and characterized. The first two classes (gamma and beta, cytoplasmic actins) were obtained from a cDNA library constructed from simian virus 40-transformed human fibroblast mRNA, and the third class (alpha, muscle actin) was obtained from a cDNA library constructed from adult human muscle mRNA. A new approach was developed to enrich for full-length cDNAs. The human fibroblast cDNA plasmid library was linearized with restriction enzymes that did not cut the inserts of interest; it was then size-fractionated on gels, and the chimeric molecules of optimal length were selected for retransformation of bacteria. When the resulting clones were screened for actin-coding sequences it was found that some full-length cDNAs were enriched as much as 50- to 100-fold relative to the original frequency of full-length clones in the total library. Two types of clones were distinguished. One of these clones encodes gamma actin and contains 100 base pairs of 5' untranslated region, the entire protein coding region, and the 3' untranslated region. The second class encodes beta actin, and the longest such clone contains 45 base pairs of 5' untranslated region plus the remainder of the mRNA extending to the polyadenylic acid tail. A third class, obtained from the human muscle cDNA library, encodes alpha actin and contains 100 base pairs of 5' untranslated region, the entire coding region, and the 3' untranslated region. Analysis of the DNA sequences of the 5' end of the clones demonstrated that although beta- and gamma-actin genes start with a methionine codon (MET-Asp-Asp-Asp and MET-Glu-Glu-Glu, respectively), the alpha-actin gene starts with a methionine codon followed by a cysteine codon (MET-CYS-Asp-Glu-Asp-Glu). Since no known actin proteins start with a cysteine, it is likely that post-translational removal of cysteine in addition to methionine accompanies alpha-actin synthesis but not beta- and gamma-actin synthesis. This observation has interesting implications both for actin function and actin gene regulation and evolution.
1,161 citations
TL;DR: The DNA sequences derived from the germ line JH-C mu region are found to be required for accurate and efficient transcription from a functionally rearranged VH promoter, and the Ig gene enhancer appears to act in a tissue-specific manner.
1,145 citations
TL;DR: These vectors have been used successfully for DNA sequencing with the dideoxy-method, and can be used for any other purpose for which M13 derivatives are used, however, the pEMBL plasmids have the advantage of being smaller than M13 vectors, and the purification of the DNA is simpler.
Abstract: We have constructed a series of plasmids, the pEMBL family, characterized by the presence of 1) the bla gene as selectable marker, 2) a short segment coding for the alpha-peptide of beta-galactosidase and containing a multiple cloning sites polylinker, 3) the intragenic region of phage F1. pEMBL plasmids have the property of being encapsidated as single stranded DNA, upon superinfection with phage F1. These vectors have been used successfully for DNA sequencing with the dideoxy-method, and can be used for any other purpose for which M13 derivatives are used. However, the pEMBL plasmids have the advantage of being smaller than M13 vectors, and the purification of the DNA is simpler. In addition, and most importantly, long inserts have a higher stability in pEMBL plasmids than M13 vectors.
TL;DR: Chimeric bacterial genes conferring resistance to aminoglycoside antibiotics have been inserted into the Agrobacterium tumefaciens tumor-inducing (Ti) plasmid and introduced into plant cells by in vitro transformation techniques.
Abstract: Chimeric bacterial genes conferring resistance to aminoglycoside antibiotics have been inserted into the Agrobacterium tumefaciens tumor-inducing (Ti) plasmid and introduced into plant cells by in vitro transformation techniques. The chimeric genes contain the nopaline synthase 5' and 3' regulatory regions joined to the genes for neomycin phosphotransferase type I or type II. The chimeric genes were cloned into an intermediate vector, pMON120, and inserted into pTiB6S3 by recombination and then introduced into petunia and tobacco cells by cocultivating A. tumefaciens cells with protoplast-derived cells. Southern hybridization was used to confirm the presence of the chimeric genes in the transformed plant tissues. Expression of the chimeric genes was determined by the ability of the transformed cells to proliferate on medium containing normally inhibitory levels of kanamycin (50 micrograms/ml) or other aminoglycoside antibiotics. Plant cells transformed by wild-type pTiB6S3 or derivatives carrying the bacterial neomycin phosphotransferase genes with their own promoters failed to grow under these conditions. The significance of these results for plant genetic engineering is discussed.
TL;DR: The usage of synonymous codons in the beta-actin gene is nonrandom, and is similar to that in the rat skeletal muscle and other vertebrate actin genes, but differs from the codon usage in yeast and soybean actin gene.
Abstract: The nucleotide sequence of the rat beta-actin gene was determined. The gene codes for a protein identical to the bovine beta-actin. It has a large intron in the 5' untranslated region 6 nucleotides upstream from the initiator ATG, and 4 introns in the coding region at codons specifying amino acids 41/42, 121/122, 267, and 327/328. Unlike the skeletal muscle actin gene and many other actin genes, the beta-actin gene lacks the codon for Cys between the initiator ATG and the codon for the N-terminal amino acid of the mature protein. The usage of synonymous codons in the beta-actin gene is nonrandom, and is similar to that in the rat skeletal muscle and other vertebrate actin genes, but differs from the codon usage in yeast and soybean actin genes.
TL;DR: Overall Conclusions are that DNA Methylation at Specific Sites is Causally Related /0 Gene Inactivation and the Drosophila Problem is a Model of Herpes Simplex Virus Latency.
Abstract: PERSPECTIVES AND SUMMARY 93 BIOCHEMISTRY OF DNA METHYLATION . . ...... ....... . . . 95 Occurrence of Modified Bases ...... .... . . . . . . ......... . . . ... .. 95 Maintenance and de novo Methylations ........ .. ... ........ ..... ..... .. ... 96 Methods to Determine Methylated Bases .. ........ .. . . . . . . . .. . . ...... . ...... ..... . .... . . . 99 Distribution of 5-Methylcytosine ... .. ... . .. .. . ....... .... . ..... .. ... .. . ... . .... ......... .... . 100 DN A Methylatiun and Structural Changes uf DN A 102 THE BIOLOGICAL FUNCTION OF DNA METHYLATION . . . . . . 103 DNA Methylation--a Regulatory Signal in Eukaryotic Gene Expression . ... . . . .. . 103 Correlations ... . ..... .. .. . . . . . . .... . .. .... . . . . . . . . . . . . ......... ......... . ......... . ...... . ...... 105 The Drosophila Problem . . .. .. ..... ..... .. .. .... . ... .... .... . . . .... ... .. . . ... .... ... .. .... . 109 Treatment of Cells with 5-Azacytidine Can Lead to the Activation of Previously Dormant Genes ..... ..... ... .. .... .. . . .... .. ..... ....... ..... . . .. ... ..... . . . .. .. . . 110 DNA Methylation at Specific Sites is Causally Related /0 Gene Inactivation ..... . . 112 Additional Observations . . .. .. . .. . . . .. .. ... . . .. . ......... . .... .... . ........ . . . . .... .... . .. . . 115 Overall Conclusions ..... 116 DNA Methylation and Genetic Recombination . .... ... . . .. .. . . .... ... .. ..... . . ..... .. .... 117 A Model of Herpes Simplex Virus Latency 117 IN SEARCH OF MORE COMPLEX GENETIC CODING SIGNALS . ......... . .. 118 OUTLOOK ........ 118
TL;DR: This chapter describes plasmids and methods for constructing fusions of any cloned gene to lacZ for study in yeast, which provide powerful tools in the analysis of the expression of yeast genes.
Abstract: Publisher Summary This chapter describes plasmids and methods for constructing fusions of any cloned gene to lacZ for study in yeast. If the gene to be fused to lacZ contains no appropriate Sau3A sites, alternative strategies to the above must be employed. If the DNA sequence of the gene to be fused to lacZ is known, inframe fusions can be made simply by choosing the appropriate gene fragment. The vector sequences that precede the yeast DNA is a yeast gene–lacZ fusion may affect gene regulation. Plasmids may be used to probe the signals that govern the initiation of transcription and translation in S. cerevisiae. These methods provide powerful tools in the analysis of the expression of yeast genes. Further, the plasmids should facilitate the expression in yeast of any cloned gene to produce the native, unfused product.
TL;DR: DNA sequence analysis of the activated oncogene from the T24 human bladder carcinoma line and two alleles of its normal cellular progenitor (c-Ha-ras-1) indicates that the genes encompass at least four exons.
Abstract: DNA sequence analysis of the activated oncogene from the T24 human bladder carcinoma line and two alleles of its normal cellular progenitor (c-Ha-ras-1) indicates that the genes encompass at least four exons, and that only a single point mutation residing within the first exon distinguishes the coding region of both alleles of the normal gene from their activated counterpart. Both versions o f the gene encode a protein which is predicted to differ from the corresponding viral gene product at three amino acid residues, one of which was previously shown to represent the major site of phosphorylation of the viral polypeptide.
TL;DR: In this article, the amplification and expression of the c-myc oncogene in a system other than B-cell lymphomas, namely human lung cancer, was reported.
Abstract: Genetic changes involving the c-myc oncogene have been observed in human tumours. In particular, the c-myc gene is translocated in Burkitt's lymphoma and is amplified in the human promyelocytic leukaemia cell line, HL-60, which contains double minute chromosomes (DMs). More recently, an amplified c-myc gene has been positioned on a chromosomal homogeneous staining region (HSR) in a human colon cancer cell line, COLO 320, with neuroendocrine properties. Furthermore, c-myc is expressed in increased amounts in some human tumour lines, and in some cases, human small cell lung cancers (SCLC) contain DMs and HSRs. These findings prompted us to study the c-myc gene and its RNA expression in a series of human lung cancer cell lines. We now report amplification and expression of the c-myc oncogene in a system other than B-cell lymphomas, namely human lung cancer. Of 18 human lung cancer cell lines tested, 8 showed an amplified 12.5-kilobase (kb) EcoRI c-myc DNA band. Of particular interest are five SCLC lines with a high degree of c-myc DNA amplification (20-76-fold) and greatly increased levels of c-myc RNA. All five lines reside in the variant class of SCLC (SCLC-V) characterized by altered morphology, lack of expression of some SCLC-differentiated functions and more malignant behaviour than pure SCLC. Three of the five lines which have been karyotyped also contain DMs or HSRs. The finding of a greatly amplified c-myc gene in all cell lines of the SCLC-V class examined strongly suggests a role for the c-myc gene in the phenotypic conversion and malignant behaviour of human lung cancer.
TL;DR: To study the regulation of the human beta-interferon (beta-IFN) gene by poly(I)-poly(C), the expression of deletion mutants of the cloned gene introduced into mouse cells on a new bovine papilloma virus vector is analyzed.
TL;DR: The plasmid-borne gene hph coding for hygromycin B phosphotransferase (HPH) in Escherichia coli has been identified and its nucleotide sequence determined.
TL;DR: The tac promoter is repressed in lacIQ strains and can be induced by isopropylthio-beta-D-galactoside (IPTG), and studies of expression of the cI repressor of bacteriophage lambda show that the tac promoters is at least five times more efficient than the lacUV5 promoter.
TL;DR: It is hypothesized that E was part of a duplicated region of over 250 nucleotides flanking the src gene in an ancestral RSV, and that differential deletion of one copy of E led to its positional difference in Pr-C and SR-A.
TL;DR: Evidence is presented that the Wx locus encodes a starch granule-bound 58 kd polypeptide that is synthesized in vitro as a 65 kd precursor and it is shown that a mutation caused by the controlling element Dissociation is attributable to an insertion of approximately 2.4 kb at the WX locus.
TL;DR: In this article, a 16-kb insert that encoded enzymatic activities for the light reaction as well as regulatory functions necessary for expression of the luminescence phenotype (Lux) was found in a clone library of hybrid plasmids containing DNA from the marine bacterium Vibrio fischeri.
TL;DR: The fusion genes were expressed in all tissues examined, but the ratio of human growth hormone messenger RNA to endogenous metallothionein-I messenger RNA varied among different tissues and different animals, suggesting that expression of the foreign genes is influenced by site of integration and tissue environment.
Abstract: The promoter or regulatory region of the mouse gene for metallothionein-I was fused to the structural gene coding for human growth hormone. These fusion genes were introduced into mice by microinjection of fertilized eggs. Twenty-three (70 percent) of the mice that stably incorporated the fusion genes showed high concentrations of human growth hormone in their serum and grew significantly larger than control mice. Synthesis of human growth hormone was induced further by cadmium or zinc, which normally induce metallothionein gene expression. Transgenic mice that expressed human growth hormone also showed increased concentrations of insulin-like growth factor I in their serum. Histology of their pituitaries suggests dysfunction of the cells that normally synthesize growth hormone. The fusion genes were expressed in all tissues examined, but the ratio of human growth hormone messenger RNA to endogenous metallothionein-I messenger RNA varied among different tissues and different animals, suggesting that expression of the foreign genes is influenced by site of integration and tissue environment.
TL;DR: DNA-mediated genetic transformation of Aspergillus nidulans has been achieved by incubating protoplasts from a strain of A. niduans carrying a deletion in the acetamidase structural gene with DNA of derivatives of plasmid pBR322 containing the cloned structural gene for acetamids.
TL;DR: An assay was developed whereby transcription of a cloned, rearranged kappa gene could be detected following its transfection into antibody-secreting mouse myeloma cells, suggesting that after rearrangement the kappa variable region promoter is activated by sequences more than 2.6 kb downstream of J kappa.
TL;DR: In vitro methods and some recently developed β -galactosidase gene fusion vectors are described, which can be used not only to measure gene expression and regulation, but also to isolate mutations and additional gene fusions.
Abstract: Publisher Summary Gene fusions can be constructed either in vivo , using spontaneous nonhomologous recombination or semi-site specific transposon recombination, or in vitro , with recombinant DNA technology. This chapter describes in vitro methods and lists some recently developed β -galactosidase gene fusion vectors. With these methods, gene-controlling elements from any source can be fused to the β -galactosidase structural gene and examined in the prokaryote bacterium Escherichia coli or the lower eukaryote yeast Saccharomyces cerevisiae. β -galactosidase gene fusions can be constructed both with transcription initiation control signals and with transcription plus translation initiation control signals. The β -galactosidase gene is convenient for making translational fusions because it is possible to remove its translation initiation region along with up to at least the first 27 amino acid codons without affecting β -galactosidase enzymic activity. The focus here is on these transcription-translation fusions because they provide all the gene initiation signals from the other gene. β -Galactosidase expression from a gene fusion can be used not only to measure gene expression and regulation, but also to isolate mutations and additional gene fusions.
Book•
01 Jan 1983
TL;DR: This sequel to The Selfish Gene argues that since genes can be said to have extended phenotypes outside the body in which they sit, the authors will have to revolutionize their views of evolutionary adaptation.
Abstract: A stimulating investigation of new perspectives on gene effects, this sequel to The Selfish Gene argues that since genes can be said to have extended phenotypes outside the body in which they sit, we will have to revolutionize our views of evolutionary adaptation. It also looks at the theory of evolutionary stable strategies, the relationship between Darwinian and Lamarckian theories of adaptation, and Dawkin's suggestion that some of the surplus DNA in eukaryotic genomes may be parasitic.
TL;DR: Results indicate that chemical carcinogenesis represents an adequate model to study the role of transforming ras genes in human neoplasia and that the twelfth codon was GAA instead of GGA of the normal allele, encoding glutamic acid in place of glycine.
Abstract: Each of nine mammary carcinomas induced by a single injection of nitroso-methylurea into 50-day-old Buf/N female rats, contained a transforming H-ras-1 gene. Molecular characterization of one of the genes revealed that the twelfth codon was GAA instead of GGA of the normal allele, encoding glutamic acid in place of glycine. These results indicate that chemical carcinogenesis represents an adequate model to study the role of transforming ras genes in human neoplasia.