Multiple positive and negative cis-acting elements mediate induced arginase (CAR1) gene expression in Saccharomyces cerevisiae.

Kovari, L. Z.; Sumrada, R; Kovari, Iulia A.; Cooper, Terrance G.

doi:10.1128/mcb.10.10.5087

Cited by 48 publications

(108 citation statements)

References 34 publications

Supporting

Mentioning

104

Contrasting

Order By: Relevance

“…We used a CYCJ-lacZ-containing vector (pNG22) described by Kovari et al (32) to assay MAG URS2 function. A 24-bp oligonucleotide containing 20 bp of the putative MAG URS2 region was cloned 3' of the CYCI UAS.…”

Section: Resultsmentioning

confidence: 99%

“…Eag I sites of pNG22 (32), which contains the CYCl UAS-lacZ fusion and a polylinker 3' to the CYCI UAS. pNG22 containing an unrelated sequence of the same size was created by digesting plasmid pNG22/MCS (15) with Eag I and selfligating the vector.…”

Section: Methodsmentioning

confidence: 99%

“…Yeast transformation was carried out as described (30,31). ,B-Galactosidase assays were as described (15,32). A 24-bp oligonucleotide containing 20 bp of the MAG URS2 region (-180 to -161; see Table 1) was cloned between the Sal I and Abbreviations: UAS, upstream activating sequence; URS, upstream repressing sequence; MeMes, methyl methanesulfonate; ssDNA, single-stranded DNA; dsDNA, double-stranded DNA.…”

Section: Methodsmentioning

confidence: 99%

See 2 more Smart Citations

Replication protein A binds to regulatory elements in yeast DNA repair and DNA metabolism genes.

Singh

Samson

1995

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

View full text Add to dashboard Cite

Saccharomyces cerevisiae responds to DNA damage by arresting cell cycle progression (thereby preventing the replication and segregation of damaged chromosomes) and by inducing the expression of numerous genes, some of which are involved in DNA repair, DNA replication, and DNA metabolism. Induction of the S. cerevisiae 3-methyladenine DNA glycosylase repair gene (MAG) by DNA-damaging agents requires one upstream activating sequence (UAS) and two upstream repressing sequences (URS1 and URS2) in the MAG promoter. Sequences similar to the MAG URS elements are present in at least 11 other S. cerevisiae DNA repair and metabolism genes. Replication protein A (Rpa) is known as a single-stranded-DNA-binding protein that is involved in the initiation and elongation steps of DNA replication, nucleotide excision repair, and homologous recombination. We now show that the MAG URS1 and URS2 elements form similar doublestranded, sequence-specific, DNA-protein complexes and that both complexes contain Rpa. Moreover, Rpa appears to bind theMAG URSi-like elements found upstream of 11 other DNA repair and DNA metabolism genes. These results lead us to hypothesize that Rpa may be involved in the regulation of a number of DNA repair and DNA metabolism genes.Alkylating agents covalently modify DNA to generate alkylated bases and are toxic, mutagenic, and carcinogenic. Some alkylated bases cause mutations because they miscode when replicated, and others cause cell death because they block DNA replication, preventing cells from proceeding properly through the cell cycle (1-3). If they are to avoid the mutagenic and cytotoxic effects of alkylating agents, cells must repair these alkylated bases before their DNA is replicated. Halting DNA replication when the genome is assaulted assures that DNA damage will be repaired before it has a chance to be encountered by the replication machinery.Saccharomyces cerevisiae, like Escherichia coli and mammalian cells, responds to DNA damage by inhibiting DNA replication and arresting cell cycle progression, thus preventing the replication of damaged chromosomes (3-6). These cells also respond by inducing the expression of numerous genes, and many of these genes are involved in DNA repair and DNA metabolism (7,8). The regulation of DNA damage-inducible regulons has been well characterized in E. coli (8-11). The SOS response is induced when single-stranded DNA (ssDNA), generated in the locale of DNA damage, activates RecA protein to facilitate the cleavage and inactivation of the LexA transcriptional repressor, derepressing a set of genes involved in DNA repair, mutagenesis, recombination, and cell division (8). Over 40 genes are regulated by OxyR and SoxRS in response to oxidative stress (9, 10), and four genes are regulated by the Ada DNA methyltransferase in response to alkylating agents (11). Methyl transfer from a methylphosphotriester lesion to the Ada Cys-69 residue converts the AdaThe publication costs of this article were defrayed in part by page charge payment. This article must theref...

show abstract

Section: Resultsmentioning

confidence: 99%

Section: Methodsmentioning

confidence: 99%

Section: Methodsmentioning

confidence: 99%

See 1 more Smart Citation

Replication protein A binds to regulatory elements in yeast DNA repair and DNA metabolism genes.

Singh

Samson

1995

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

View full text Add to dashboard Cite

show abstract

“…The high levels of CAR1 mRNA likely derived from two sources. First, the CAR1 promoter contains multiple demonstrably functional cis-acting elements (12 sequences), including three CAR1 UAS I elements that mediate strong arginineinduced CAR1 transcription and one that is homologous to a Gln3/Gat1 binding site (Kovari et al 1990(Kovari et al , 1993aViljoen et al 1992). The vacuole contains millimolar levels of arginine that are mobilized during starvation; this induces arginase (CAR1) production, accounting for the high levels of CAR1 expression observed (Wiemken et al 1970;Wiemken and Durr 1974;Zacharski and Cooper 1978;Sumrada and Cooper 1978).…”

Section: Relationship Of Nuclear Gln3 and Gat1 Localization To Cell Cmentioning

confidence: 99%

Nitrogen Starvation and TorC1 Inhibition Differentially Affect Nuclear Localization of the Gln3 and Gat1 Transcription Factors Through the Rare Glutamine tRNACUG in Saccharomyces cerevisiae

2014

View full text Add to dashboard Cite

A leucine, leucyl-tRNA synthetase-dependent pathway activates TorC1 kinase and its downstream stimulation of protein synthesis, a major nitrogen consumer. We previously demonstrated, however, that control of Gln3, a transcription activator of catabolic genes whose products generate the nitrogenous precursors for protein synthesis, is not subject to leucine-dependent TorC1 activation. This led us to conclude that excess nitrogen-dependent down-regulation of Gln3 occurs via a second mechanism that is independent of leucine-dependent TorC1 activation. A major site of Gln3 and Gat1 (another GATA-binding transcription activator) control occurs at their access to the nucleus. In excess nitrogen, Gln3 and Gat1 are sequestered in the cytoplasm in a Ure2-dependent manner. They become nuclear and activate transcription when nitrogen becomes limiting. Long-term nitrogen starvation and treatment of cells with the glutamine synthetase inhibitor methionine sulfoximine (Msx) also elicit nuclear Gln3 localization. The sensitivity of Gln3 localization to glutamine and inhibition of glutamine synthesis prompted us to investigate the effects of a glutamine tRNA mutation (sup70-65) on nitrogen-responsive control of Gln3 and Gat1. We found that nuclear Gln3 localization elicited by short-and long-term nitrogen starvation; growth in a poor, derepressive medium; Msx or rapamycin treatment; or ure2D mutation is abolished in a sup70-65 mutant. However, nuclear Gat1 localization, which also exhibits a glutamine tRNA CUG requirement for its response to short-term nitrogen starvation or growth in proline medium or a ure2D mutation, does not require tRNA CUG for its response to rapamycin. Also, in contrast with Gln3, Gat1 localization does not respond to long-term nitrogen starvation. These observations demonstrate the existence of a specific nitrogen-responsive component participating in the control of Gln3 and Gat1 localization and their downstream production of nitrogenous precursors. This component is highly sensitive to the function of the rare glutamine tRNA CUG , which cannot be replaced by the predominant glutamine tRNA CAA . Our observations also demonstrate distinct mechanistic differences between the responses of Gln3 and Gat1 to rapamycin inhibition of TorC1 and nitrogen starvation.

show abstract

“…The CAR1 upstream regulatory region contains four discrete cis-acting elements: two inducer-independent upstream activation sequences (UASs), UASCl and UASC2; an inducer-dependent UAS, UAS,; and an upstream repression sequence, URS1 (11,13,15,23,[28][29][30][31]. The URS1 site is found upstream of many yeast genes and has been shown to bind a heteromeric protein (14,15,30).…”

mentioning

confidence: 99%

Participation of RAP1 protein in expression of the Saccharomyces cerevisiae arginase (CAR1) gene

Kovari

Cooper

1993

J Bacteriol

Self Cite

View full text Add to dashboard Cite

Regulated expression of the inducible arginase (CARl) gene of Saccharomyces cerevisiae has been shown to require (2, 6, 19-21, 27b). However, it was recently shown that only arginine transport is NCR sensitive; the apparent NCR sensitivity of CARl expression is in reality NCR-sensitive inducer exclusion (7).The CAR1 upstream regulatory region contains four discrete cis-acting elements: two inducer-independent upstream activation sequences (UASs), UASCl and UASC2; an inducer-dependent UAS, UAS,; and an upstream repression sequence, URS1 (11,13,15,23,[28][29][30][31]. The URS1 site is found upstream of many yeast genes and has been shown to bind a heteromeric protein (14,15,30). The functional integration of these elements and the proteins associated with them is proposed to mediate controlled CARl expression (30). In the absence of inducer, UASC1 and UASc and their associated proteins are capable of activating transcription but are prevented from doing so by the strong negative action of URS1 and its associated proteins. When arginine is added, UASJ becomes functional, the combined action of all three UAS elements operating simultaneously overcomes the negative action of URS1, and expression occurs (13).For an understanding of the biochemical mechanisms through which controlled expression is accomplished, it is necessary to identify all the proteins that interact with the CAR] cis-acting elements. We recently reported that UASC1 and UASC2 contained ABF-1 binding sites (11). Both sites were required for the support of wild-type levels of transcriptional activation. However, one of the sites was significantly more important than the other.Work from several other laboratories suggested that the ABF-1 transcription factor often functions in association with a second factor, RAP1 (3-5, 16, 27 303 (25). The RAP1 DNA binding domain is located between residues 361 and 596 of the 827-amino-acid protein (9). Therefore, the truncated protein retains the DNA binding site. Extracts from this strain produce a higher-mobility band shift of HIS4 upstream sequences than extracts from the wild-type strain do (25).The Escherichia coli strain used for cloning was HB101 (hsd-20 levB supE44 ara-14 galK2 lacY] proA2 rpsL20xyl-5 mtl-1 recA13 mcrB). Yeast

show abstract

Multiple positive and negative cis-acting elements mediate induced arginase (CAR1) gene expression in Saccharomyces cerevisiae.

Cited by 48 publications

References 34 publications

Replication protein A binds to regulatory elements in yeast DNA repair and DNA metabolism genes.

Replication protein A binds to regulatory elements in yeast DNA repair and DNA metabolism genes.

Nitrogen Starvation and TorC1 Inhibition Differentially Affect Nuclear Localization of the Gln3 and Gat1 Transcription Factors Through the Rare Glutamine tRNACUG in Saccharomyces cerevisiae

Participation of RAP1 protein in expression of the Saccharomyces cerevisiae arginase (CAR1) gene

Contact Info

Product

Resources

About