cis-Dominant mutations which dramatically enhance DUR1,2 gene expression without affecting its normal regulation.

Chisholm, G E; Cooper, Terrance G.

doi:10.1128/mcb.4.5.947

Cited by 12 publications

(14 citation statements)

References 33 publications

(11 reference statements)

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“…First, we worked only with the poly(A)+ RNA fraction from cells growing exponentially in rich, glucose-containing medium under aerobic conditions. Thus, any regions producing only nonpolyadenylated transcripts (20,36), including any tRNA genes, probably would have been overlooked, as would any genes that are transcribed appreciably only in other cell types or under other growth conditions (11,38,50,68,76). Second, there are some residual uncertainties in our results that might reflect the presence of additional transcribed regions (see Results) (see also Fig.…”

Section: Discussionmentioning

confidence: 62%

“…Similar numbers have been obtained with other cloned yeast chromosome segments in which the transcribed regions have been mapped. Taken together, the CDC24-PYKJ region (this study), the histone H2A and H2B gene regions (21), several ribosomal protein gene regions (33,77), the HIS3 region (70), the GAL7,10,1 region (68), the CYCJ-CDC8 region (4,16,32,38,60,62; L. Melnick, personal communication), the CDC36 and CDC37 regions (7), the ACTJ-TUB2 region (17,72), the RNA3 and RNAJJ regions (34), the DURI,2-MET8 region (11,18), the DAL gene region (79), the CDC10-CEN3 region (28,78), the SIR2, SIR3, and SIR4 regions (25,63), and the MET14-CENJI region (78) (20). On the assumption that the average S. cerevisiae transcript is 1.5 kb, this yielded an estimate of 3,000 to 4,000 different transcribed regions (20).…”

Section: Discussionmentioning

confidence: 99%

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Molecular cloning of chromosome I DNA from Saccharomyces cerevisiae: isolation and characterization of the CDC24 gene and adjacent regions of the chromosome.

Coleman¹,

Steensma²,

Kaback³

et al. 1986

Mol. Cell. Biol.

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Molecular cloning techniques were used to isolate and characterize the DNA including and surrounding the CDC24 and PYKI genes on the left arm of chromosome I of the yeast Saccharomyces cerevisiae. A plasmid that complemented a temperature-sensitive cdc24 mutation was isolated from a yeast genomic DNA library in a shuttle vector. Plasmids containing pykl-complementing DNA were obtained from other investigators. Several lines of evidence (including one-step gene replacement experiments) demonstrated that the complementing plasmids contained the bona fide CDC24 and PYKI genes. These sequences were then used to isolate additional DNA from chromosome I by probing a yeast genomic DNA library in a lambda vector. A total of 28 kilobases (kb) of contiguous DNA surrounding the CDC24 and PYKI genes was isolated, and a restriction map was determined. Electron microscopy of R-loop-containing DNA and RNA blot hybridization analyses indicated that an 18-kb segment contained at least seven transcribed regions, only three of which corresponded to previously known genes (CDC24, PYKI, and CYC3). Southern blot hybridization experiments suggested that none of the genes in this region was duplicated elsewhere in the yeast genome. The centers of CDC24 and PYKI were only -7.5 kb apart, although the genetic map distance between them is -13 centimorgans. As previous studies with S. cerevisiae have indicated that 1 centimorgan generally corresponds to -3 kb, the region between CDC24 and PYKI appears to undergo meiotic recombination at an unusually high frequency.

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Section: Discussionmentioning

confidence: 62%

Section: Discussionmentioning

confidence: 99%

Molecular cloning of chromosome I DNA from Saccharomyces cerevisiae: isolation and characterization of the CDC24 gene and adjacent regions of the chromosome.

Coleman¹,

Steensma²,

Kaback³

et al. 1986

Mol. Cell. Biol.

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“…The phenotype of DUR80 mutant strains, which is expressed both in haploid and MATa/MATa diploids, does not appear to be an alteration of the control system for this gene because its expression remained fully inducible and sensitive to nitrogen repression. Rather, we found much higher levels of DURI,2-specific RNA under both induced and uninduced conditions, i.e., enhanced expression appeared to be superimposed on normal regulation of the gene (9). The second class of mutants, isolated both in our laboratory and in that of Wiame, is designated DUR1,2-oh.…”

mentioning

confidence: 91%

“…In sum, the steady-state levels of DUR1,2 RNA qualitatively matched those reported earlier for urea amidolyase activity and protein concentration (4,8,25,37). In addition to the dal80 and dal81 mutant loci described above, two new classes of cis-dominant mutations have been isolated (9,11,27). The first class of mutations was designated DUR80.…”

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confidence: 99%

Induction and repression of the urea amidolyase gene in Saccharomyces cerevisiae.

Genbauffe¹,

Cooper²

1986

Mol. Cell. Biol.

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The DUR1,2 gene from Saccharomyces cerevisiae has been isolated on recombinant plasmids along with all DNA between the DURI,2 and MET8 loci. DUR1,2 was found to encode a 5.7-kilobase transcript, which is consistent with our earlier suggestion that the DURi and DUR2 loci are two domains of a single multifunctional gene. Steady-state levels of the DUR1,2 transcript responded to induction and nitrogen catabolite repression in the same way as urea amidolyase activity. da)81 mutants (grown with inducer) contained barely detectable amounts of DURI,2 RNA, whereas dal80 mutants (grown without inducer) contained the same amount as a wild-type induced culture. These observations support our earlier hypothesis that DUR1,2 is transcriptionally regulated, with control being mediated by the DAL80 and DAL8I gene products. We cloned the DUR1,2_Oh mutation and found it to be a Ty insertion near sequences required for complementation of durl,2 mutations.The ROAM phenotype of the DUR1,2_Oh mutation is sharply different from that of cis-dominant, DUR80 mutations, which enhance DUR1,2 expression but do not affect the normal control pattern of the gene. There is evidence that DUR80 mutations may also be Ty insertions, which generate phenotypes that are different from those in DUR1,2-Oh mutations.An appreciation for the molecular mechanisms involved in control and integration of procaryotic metabolic pathways has been gained by studying regulons with widely differing physiological functions (28, 31). Similar information is now beginning to accumulate for eucaryotic systems (42). Nitrogen catabolic systems are particularly useful for such investigations, because most are subject to multiple layers of regulation. Genes encoding the allantoin-degradative system in Saccharomyces cerevisiae, for example, respond to both induction and nitrogen catabolite repression (15,25,38 its 3-min half-life after the addition of asparagine to the medium (12). In contrast, enzyme activity is not lost after the addition of a repressive nitrogen source, but continued enzyme synthesis ceases (12). The lack of enzyme induction does not result from inducer exclusion, because daI80 mutants, which do not require the presence of an inducer for enzyme production, are similarly devoid of allantoindegrading enzymes when grown in glucose-asparagine medium (8).Earlier kinetic studies are consistent with the hypothesis that both induction and nitrogen catabolite repression are exerted at gene expression (1-4, 16, 24-26

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“…All three DUR80 alleles were also Ty insertions between -644 and -653 immediately 5' of the native DUR1,2 UASs. We suggest that the difference in DURJ,2-0h and DUR80 phenotypes depends on whether the native cis-acting elements and transcription factors associated with them can operate.…”

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confidence: 99%

Ty insertions upstream and downstream of native DUR1,2 promoter elements generate different patterns of DUR1,2 expression in Saccharomyces cerevisiae

Chisholm

Cooper

1992

J Bacteriol

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Expression of allantoin pathway genes is subject to induction and nitrogen catabolite repression. Two classes of cis-dominant mutations (DUR80 and DUR1,2-Oh) result in overproduction of DURI,2 mRNA. In DUR80 mutants, DURJ,2 expression remained inducible, nitrogen catabolite repression sensitive, and unresponsive to cell ploidy, i.e., overproduction was superimposed on normal gene regulation. DUR1,2-0h mutations, in contrast, generated a pattern of DUR1,2 expression similar to that often reported when a Ty element inserts upstream of a gene, the ROAM phenotype. We analyzed four independent DUR80 and DUR1,2-0h alleles. The DURJ,2-0Y mutation was, as expected, a Ty insertion at -445 3' of the native DUR1,2 upstream activation sequences (UASs). All three DUR80 alleles were also Ty insertions between -644 and -653 immediately 5' of the native DUR1,2 UASs. We suggest that the difference in DURJ,2-0h and DUR80 phenotypes depends on whether the native cis-acting elements and transcription factors associated with them can operate. If they can, enhancement of normally regulated DURJ,2 expression is observed. This is a novel phenotype for Ty insertions. If the native DURJ,2 cis-acting elements are not present, the case when Ty insertion occurs 3' of them, a ROAM phenotype is generated. Nitrogen-regulated upstream activation sequence (UASNTR)-homologous sequences present in the Ty 8 elements rather than cis-acting elements required for Ty transcription are the most likely candidates to serve as the cis-acting elements mediating the DUR80 phenotype.

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cis-Dominant mutations which dramatically enhance DUR1,2 gene expression without affecting its normal regulation.

Cited by 12 publications

References 33 publications

Molecular cloning of chromosome I DNA from Saccharomyces cerevisiae: isolation and characterization of the CDC24 gene and adjacent regions of the chromosome.

Molecular cloning of chromosome I DNA from Saccharomyces cerevisiae: isolation and characterization of the CDC24 gene and adjacent regions of the chromosome.

Induction and repression of the urea amidolyase gene in Saccharomyces cerevisiae.

Ty insertions upstream and downstream of native DUR1,2 promoter elements generate different patterns of DUR1,2 expression in Saccharomyces cerevisiae

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