Duplicate upstream activating sequences in the promoter region of the Saccharomyces cerevisiae GAL7 gene

Tajima, M; Nogi, Yasuhisa; Fukasawa, Toshio

doi:10.1128/mcb.6.1.246-256.1986

Cited by 5 publications

(2 citation statements)

References 52 publications

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“…The screen of nuclear DNA sequences was originally motivated by our observation that some nuclear genes contain mitochondrial RNA polymerase promoters functional in in vitro assays (48). These nuclear DNA sequences were useful as a collection of critical promoters but were also interesting because, in some cases, the mitochondrial promoter sequence was coincident with the TATA box for the nuclear RNA polymerase II transcription apparatus (38,43). In particular, the coordinately regulated GAL7.…”

Section: Discussionmentioning

confidence: 99%

“…higher levels of HIS3 gene expression (41). To test the effectiveness of these plate assays, the strains were also transformed with a YCp19 plasmid bearing the HIS3 gene with a fully functional RNA polymerase II promoter (YCp19-HIS3+) and a YCp19 plasmid bearing the HIS3 gene lacking all promoter elements (YCp19-his3-) (30,43). Previous Northern blot experiments have demonstrated that YCpl9-HIS3+ produces a strong HIS3 band corresponding to an estimated 5 to 10 mRNA molecules per cell, whereas YCpl9-his3produces very low levels of HIS3 mRNA (28).…”

Section: Fig 3 (A)mentioning

confidence: 99%

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Use of Yeast Nuclear DNA Sequences To Define the Mitochondrial RNA Polymerase Promoter In Vitro

Marczynski

Schultz

Jaehning

1989

Molecular and Cellular Biology

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We have extended an earlier observation that the TATA box for the nuclear GAL10 gene serves as a promoter for the mitochondrial RNA polymerase in in vitro transcription reactions (C. S. Winkley, M. J. Keller, and J. A. Jaehning, J. Biol. Chem. 260:14214-14223, 1985). In this work, we demonstrate that other nuclear genes also have upstream sequences that function in vitro as mitochondrial RNA polymerase promoters. These genes include the GAL7 and MEL1 genes, which are regulated in concert with the GAL10 gene, the sigma repetitive element, and the 2 microns plasmid origin of replication. We used in vitro transcription reactions to test a large number of nuclear DNA sequences that contain critical mitochondrial promoter sequences as defined by Biswas et al. (T. K. Biswas, J. C. Edwards, M. Rabinowitz, and G. S. Getz, J. Biol. Chem. 262:13690-13696, 1987). The results of these experiments allowed us to extend the definition of essential promoter elements. This extended sequence, -ACTATAAACGatcATAG-, was frequently found in the upstream regulatory regions of nuclear genes. On the basis of these observations, we hypothesized that either (i) a catalytic RNA polymerase related to the mitochondrial enzyme functions in the nucleus of the yeast cell or (ii) a DNA sequence recognition factor is shared by the two genetic compartments. By using cells deficient in the catalytic core of the mitochondrial RNA polymerase (rpo41-) and sensitive assays for transcripts initiating from the nuclear promoter sequences, we have conclusively ruled out a role for the catalytic RNA polymerase in synthesizing transcripts from all of the nuclear sequences analyzed. The possibility that a DNA sequence recognition factor functions in both the nucleus and the mitochondria remains to be tested.

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Section: Discussionmentioning

confidence: 99%

Section: Fig 3 (A)mentioning

confidence: 99%

Use of Yeast Nuclear DNA Sequences To Define the Mitochondrial RNA Polymerase Promoter In Vitro

Marczynski

Schultz

Jaehning

1989

Molecular and Cellular Biology

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Balancing transcriptional interference and initiation on the GAL7 promoter of Saccharomyces cerevisiae

Greger

Aranda

Proudfoot

2000

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

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Transcriptional termination of the GAL10 gene in Saccharomyces cerevisiae depends on the efficiency of polyadenylation. Either cis mutations in the poly(A) signal or trans mutations of mRNA 3 end cleavage factors result in GAL10 read-through transcripts into the adjacent GAL7 gene and inactivation (occlusion) of the GAL7 promoter. Herein, we present a molecular explanation of this transcriptional interference phenomenon. In vivo footprinting data reveal that GAL7 promoter occlusion is associated with the displacement of Gal4p transcription factors from the promoter. Interestingly, overexpression of Gal4p restores promoter occupancy, activates GAL7 expression, and rescues growth on the otherwise toxic galactose substrate. Our data therefore demonstrate a precise balance between transcriptional interference and initiation.T ranscriptional termination of RNA polymerase II (Pol II) is linked to mRNA 3Ј processing, which is a two-step reaction consisting of endonucleolytic cleavage of the RNA precursor and subsequent polyadenylation (1). Mutation of poly(A) signals results in increased transcription beyond the poly(A) site of a gene (1) and recently has been shown to depend on the activity of 3Ј end processing factors (2). Some RNA processing factors are associated with elongating Pol II, indicating a tight link between transcription and RNA processing (3, 4). In contrast to Pol I (5) and Pol III, termination of Pol II occurs at variable, ill-defined positions downstream of the poly(A) site of a gene (1). These findings suggest that transcriptional termination can be a random process.In some instances, however, termination of transcription must occur efficiently, because enhanced transcriptional read-through can result in inhibition of an adjacent, downstream promoter and also perturbs origin of replication and centromere function (6-9). Closely spaced genes are particularly prone to promoter occlusion, especially when they are expressed at the same time, as in the case of the GAL10 and GAL7 genes of Saccharomyces cerevisiae. These two genes are separated by 600 bp and are activated simultaneously to high levels by galactose (gal). We have shown previously that deletion of the GAL10 poly(A) signal leads to enhanced GAL10 read-through transcription and formation of GAL10-7 bicistronic transcripts, which in turn results in inhibition of the downstream GAL7 promoter (ref. 10; Fig. 1A). Gal7p (gal uridyl transferase) catalyzes conversion of the metabolic intermediate gal 1-phosphate, which is otherwise toxic. Inhibition of Gal7p expression by transcriptional readthrough into the GAL7 promoter renders cells gal sensitive and results in a Gal Ϫ phenotype, emphasizing the importance of termination in this system.Transcription of the GAL genes depends on the transcription factor Gal4p, which binds to 17-bp sequence elements within the promoter regions (11, 12). The binding affinity of Gal4p for these sites depends on both the sequence and spacing between the critical outer nucleotide triplets (CGG. . . GGC), which are conta...

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Structural Alterations of the Nucleolus in Mutants of Saccharomyces cerevisiae Defective in RNA Polymerase I

Oakes

Nogi

Clark

et al. 1993

Molecular and Cellular Biology

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We have previously constructed mutants of Saccharomyces cerevisiae in which the gene for the second-largest subunit of RNA polymerase I (Pol I) is deleted. In these mutants, rRNA is synthesized by RNA polymerase II from a hybrid gene consisting of the 35S rRNA coding region fused to the GAL7 promoter on a plasmid. These strains thus grow in galactose but not glucose media. By immunofluorescence microscopy using antibodies against the known nucleolar proteins SSB1 and fibrillarin, we found that the intact crescent-shaped nucleolar structure is absent in these mutants; instead, several granules (called mininucleolar bodies [MNBs]) that stained with these antibodies were seen in the nucleus. Conversion of the intact nucleolar structure to MNBs was also observed in Pol I temperature-sensitive mutants at nonpermissive temperatures. These MNBs may structurally resemble prenucleolar bodies observed in higher eukaryotic cells and may represent a constituent of the normal nucleolus. Furthermore, cells under certain conditions that inhibit rRNA synthesis did not cause conversion of the nucleolus to MNBs. Thus, the role of Pol I in the maintenance of the intact nucleolar structure might include a role as a structural element in addition to (or instead of) a functional role to produce rRNA transcripts. Our study also shows that the intact nucleolar structure is not absolutely required for rRNA processing, ribosome assembly, or cell growth and that MNBs are possibly functional in rRNA processing in the Pol I deletion mutants.

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Duplicate upstream activating sequences in the promoter region of the Saccharomyces cerevisiae GAL7 gene

Cited by 5 publications

References 52 publications

Use of Yeast Nuclear DNA Sequences To Define the Mitochondrial RNA Polymerase Promoter In Vitro

Use of Yeast Nuclear DNA Sequences To Define the Mitochondrial RNA Polymerase Promoter In Vitro

Balancing transcriptional interference and initiation on the GAL7 promoter of Saccharomyces cerevisiae

Structural Alterations of the Nucleolus in Mutants of Saccharomyces cerevisiae Defective in RNA Polymerase I

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