Genes for the 70,000 dalton heat shock protein in two cloned D. melanogaster DNA segments

Artavanis‐Tsakonas, Spyros; Schedl, Paul; Mirault, Marc-Édouard; Moran, Larry A.; Lis, John T.

doi:10.1016/0092-8674(79)90290-3

Cited by 99 publications

(31 citation statements)

References 22 publications

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“…Recently, it has been found that the 3' end of mouse Ul snRNA forms efficiently only when it is located within 400 nt of the promoter (55a). As the distance from the promoter increases from 200 to 500 nt, the amount of proper 3 (68,69). They find that the arrested polymerases can be released by Sarkosyl.…”

Section: Discussionmentioning

confidence: 99%

Control of formation of two distinct classes of RNA polymerase II elongation complexes.

1992

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We have examined elongation by RNA polymerase II initiated at a promoter and have identified two classes of elongation complexes. Following initiation at a promoter, all polymerase molecules enter an abortive mode of elongation. Abortive elongation is characterized by the rapid generation of short transcripts due to pausing of the polymerase followed by termination of transcription. Termination of the early elongation complexes can be suppressed by the addition of 250 mM KCI or 1 mg of heparin per ml soon after initiation. Elongation complexes of the second class carry out productive elongation in which long transcripts can be synthesized. Productive elongation complexes are derived from early paused elongation complexes by the action of a factor which we call P-TEF (positive transcription elongation factor). P-TEF is inhibited by 5,6-dichloro-1-B-Dribofuranosylbenzimidazole at concentrations which have no effect on the initiation of transcription. By using templates immobilized on paramagnetic particles, we show that isolated preinitiation complexes lack P-TEF and give rise to transcription complexes which can carry out only abortive elongation. The ability to carry out productive elongation can be restored to isolated transcription complexes by the addition of P-TEF after initiation. A model is presented which describes the role of elongation factors in the formation and maintenance of elongation complexes. The model is consistent with the available in vivo data concerning control of elongation and is used to predict the outcome of other potential in vitro and in vivo experiments.It is now clear that the transcription of eucaryotic genes is controlled during the elongation phase as well as at initiation. The number of genes for which elongational control has been implicated is growing (80). The three proto-oncogenes c-myc (52, 61, 65, 79, 85), c-myb (4, 62), and c-fos (16, 70) have been shown to be controlled at elongation. Adenovirus (37,66,73), simian virus 40 (36, 67), minute virus of mice (39), and human immunodeficiency virus (HIV) (40,41,75,83) have been demonstrated to have specific blocks to transcription elongation. The mRNA levels for the adenosine deaminase genes of humans and mice are at least partly controlled by a regulated block to elongation (13,14,42,48,58). Many genes in Drosophila melanogaster have RNA polymerase II molecules arrested early after initiation (68,69). While control of elongation has been implicated in these examples, very little is known about the molecular mechanisms involved.A number of factors that influence elongation and termination by procaryotic RNA polymerase have been defined (86). In particular, the N and Q protein-mediated antitermination systems of lambda and similar bacteriophages provide a model for how specific gene expression can be controlled by modifying RNA polymerase elongation. The mechanism by which lambda Q protein functions has been partially elucidated (87). A key feature in this process is the pausing of the RNA polymerase downstream of the initiation...

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Section: Discussionmentioning

confidence: 99%

Control of formation of two distinct classes of RNA polymerase II elongation complexes.

1992

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“…The heavy line shows D. inelanogaster DNA; the dashed line shows DNA which in S. cerevisiae Y70 is pBR322 DNA. Hybridization was with the purified BamHI-SalI fragment from a cloned 87A copy of hsp70 (1), which hybridizes with a 2,755-bp BamHIEcoRI restriction fragment in S. cerevisiae Y70 and 9.5-, 8-, 3.2-, 2.25-, and 1.6-kb restriction fragments in D. melanogaster. Hybridization to the larger fragments is not shown.…”

Section: Resultsmentioning

confidence: 99%

High-resolution mapping of DNase I-hypersensitive sites of Drosophila heat shock genes in Drosophila melanogaster and Saccharomyces cerevisiae.

Costlow¹,

Lis²

1984

Mol. Cell. Biol.

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High-resolution analysis of the chromatin structure of the promoter regions of five Drosophila heat shock genes showed a similar location for the hypersensitive sequences relative to the start of transcription. For each of the five genes examined-those coding for hsp27, hsp26, hsp23, hsp70, and hsp83-the DNase Ihypersensitive sites in Drosophila melanogaster nuclei mapped to two regions upstream of the coding region. These sites occurred on the average, 115 and 17 base pairs upstream from the start of transcription of the five heat shock genes examined. This latter site corresponded to sequences at or near the TATA consensus sequence. Sites even further upstream of the hsp27, hsp26, and hsp83 genes were also evident. Additionally, for the two genes examined-hsp70 and hsp83-the DNase I-hypersensitive sites were preserved, at least within this level of resolution (± 10 base pairs), when the Drosophila genes were integrated into the Saccharomyces cerevisiae genome. This result indicates that the signals responsible for generating these hypersensitive sites are inherent in the DNA sequences and, in this case, are not highly species specific.The ability of a eucaryotic gene to be transcribed in vivo has been correlated with discontinuities in the chromatin structure upstream of that gene. These discontinuities have been detected as containing DNA sequences that are extremely susceptible to double-stranded cleavage by DNase I, as well as by other types of nucleases (43). The accumulated evidence suggests that these structures, called DNase Ihypersensitive sites, are necessary but insufficient for eucaryotic gene expression (10). These structures may be a consequence of either a specific DNA configuration that prevents normal nucleosome structure or a specific protein-DNA interaction or both.The relationship between the existence of DNase I-hypersensitive sites upstream of eucaryotic genes and the expression of those genes is correlative at present. Neither the molecular composition nor the function of these regions is clear. Examination and comparison of DNA sequences surrounding and composing DNase I-hypersensitive sites should help to clarify the involvement of the DNA sequence in their generation. Yet, to date, in only a few studies (13,25,31,33) have hypersensitive sites been mapped with a resolution high enough that the DNA sequence itself could be correlated with their locations.The Drosophila heat shock genes are members of a family of coordinately regulated genes. Heat shock induction of these genes occurs in all tissues examined, including Drosophila melanogaster tissue culture cells, upon a temperature shift from 23 to 37°C (2). Induction of subsets of the heat shock gene family at different developmental stages has also been reported (34, 44). DNase I-hypersensitive sites upstream of heat shock genes have been mapped previously with a resolution of ca. +50 to 100 base pairs (bp) (20,42,43). The sequences upstream of these genes are hypersensitive to nucleases even at 23°C, which may be a reflection either o...

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“…The major, 70,000-dalton, heat-inducible protein (hsp7o) in D. melanogaster is encoded at two cytogenetic loci, 87A7 and 87C1 (19)(20)(21)(22). The coding sequence for the hsp70 mRNA is 2.1 kilobases (kb) long, contains no introns, and is duplicated at both loci.…”

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confidence: 99%

“…The coding sequence for the hsp70 mRNA is 2.1 kilobases (kb) long, contains no introns, and is duplicated at both loci. The duplicated sequences ("z element") include 400 bases 5' to the transcribed region which is conserved in all copies (20,(23)(24)(25). At 87A7, two z elements are arranged in a diverging orientation separated by a 1.2-kb spacer region (26,27).…”

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confidence: 99%

Variation at the 87A heat shock locus in Drosophila melanogaster

Brown¹

1983

Proc. Natl. Acad. Sci. U.S.A.

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Restriction maps for 25 kilobases of DNA around the 87A7 heat shock locus have been determined in 29 chromosomes isolated from a natural population. The heterozygosity per nucleotide and the proportion of polymorphic nucleotide sites were estimated to be 0.0024 and 0.007, respectively. The mean number of insertional differences in this region between random pairs of chromosomes was 0.95. A significant amount of this variation was due to the insertion of large transposable elements. All the insertion/deletion events were found in a region less than 2 kilobases in size. This could either be due to nonrandom integration or to differences in the intensity of selection against DNA insertion at different sites.Several estimates of the level of variation in mitochondrial DNA from natural populations have now been made (1)(2)(3)(4)(5). This work has generally involved mapping restriction site polymorphisms and various approaches to the analysis of such data have now been published (6-10). The study of DNA-level variation in nuclear genes is technically more complex and to date three systematic population surveys have been completed-one on the ,B-globin region in man (11) and two on the alcohol dehydrogenase (Adh) locus in Drosophila melanogaster (12, 13). Much information has been collected at other loci in man (14-18) but, in general, those studies were conditioned by the prior discovery of variation and do not provide an unbiased estimate of its overall level. In this paper I present the results of a survey on variation in restriction map at the 87A7 heat shock locus in a single natural population of D. melanogaster.The major, 70,000-dalton, heat-inducible protein (hsp7o) in D. melanogaster is encoded at two cytogenetic loci, 87A7 and 87C1 (19-22). The coding sequence for the hsp70 mRNA is 2.1 kilobases (kb) long, contains no introns, and is duplicated at both loci. The duplicated sequences ("z element") include 400 bases 5' to the transcribed region which is conserved in all copies (20,(23)(24)(25). At 87A7, two z elements are arranged in a diverging orientation separated by a 1.2-kb spacer region (26,27 (28).In this paper I present data that indicate that the conservation of gene arrangement observed in this species subgroup has been achieved in spite of high levels of intraspecific insertion/deletion variation. Although such events may occur outside the regions required for correct expression of heat shock sequences (29, 30), they nevertheless appear to affect fitness sufficiently for them to be screened out by natural selection. MATERIALS AND METHODSTwenty-nine D. melanogaster third chromosomes were obtained from a large collection made in June 1977 from a natural population in Raleigh, N.C. Since that time the chromosomes have been maintained over In (3LR)TM6 Ubx. Methods used for setting up these lines have been described elsewhere (31). Before extracting DNA, the 87A region of each wild-derived chromosome was uncovered by crossing the balanced stock to a deficiency stock DJ(3R)karD3 and selecting +/de...

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Genes for the 70,000 dalton heat shock protein in two cloned D. melanogaster DNA segments

Cited by 99 publications

References 22 publications

Control of formation of two distinct classes of RNA polymerase II elongation complexes.

Control of formation of two distinct classes of RNA polymerase II elongation complexes.

High-resolution mapping of DNase I-hypersensitive sites of Drosophila heat shock genes in Drosophila melanogaster and Saccharomyces cerevisiae.

Variation at the 87A heat shock locus in Drosophila melanogaster

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