The role of RAP1 in the regulation of the MAT alpha locus
- PMID: 1990267
- PMCID: PMC359781
- DOI: 10.1128/mcb.11.2.1069-1079.1991
The role of RAP1 in the regulation of the MAT alpha locus
Abstract
The RAP1 gene of Saccharomyces cerevisiae encodes an abundant DNA-binding protein, also known as GRF1, TBA, or TUF, that binds to many sites in the yeast genome in vitro. These sites define a consensus sequence, [sequence: see text], and deletion analyses of genes that contain this sequence have implicated the involvement of RAP1 in numerous cellular processes, including gene activation and repression. The MAT alpha locus, required for determination of the alpha cell type in yeast cells, contains a RAP1 binding site; this site coincides with the MAT alpha upstream activating sequence (UAS) and is necessary for expression of the two genes encoded by the MAT alpha locus, MAT alpha 1 and MAT alpha 2. We show that the MAT alpha UAS is sufficient to activate transcription from a promoterless gene fusion of the yeast CYC1 upstream region and the lacZ gene. Constructs containing only the MAT alpha UAS generated elevated levels of beta-galactosidase activity which were indistinguishable from those of constructs containing the entire MAT alpha intergenic region. Further, the MAT alpha UAS has an intrinsic polarity of transcriptional activation; transcription of CYC1-lacZ was six- to sevenfold higher when the UAS was oriented in the direction normally associated with MAT alpha 2 transcription. Point mutations in the MAT alpha UAS that reduce MAT alpha expression three- to fivefold resulted in a bi-mating phenotype, while a mutation that reduced MAT alpha expression still further resulted in an a-mating phenotype. We isolated plasmids from a high-copy-number yeast library that suppressed the bi-mating defect of point mutations in the MAT alpha UAS, and the most effective dosage suppressor contained the gene encoding RAP1. A temperature-sensitive rap1 mutant bi-mates at the semipermissive temperature. Double mutants at rap1 and mat alpha mate exclusively as a cells, at all temperatures, and do not express detectable levels of MAT alpha RNA. These data provide evidence that the RAP1 gene product functions at the MAT alpha UAS in vivo.
Similar articles
-
RAP1 protein activates and silences transcription of mating-type genes in yeast.Genes Dev. 1991 Apr;5(4):616-28. doi: 10.1101/gad.5.4.616. Genes Dev. 1991. PMID: 2010087
-
Multiple factors bind the upstream activation sites of the yeast enolase genes ENO1 and ENO2: ABFI protein, like repressor activator protein RAP1, binds cis-acting sequences which modulate repression or activation of transcription.Mol Cell Biol. 1990 Sep;10(9):4872-85. doi: 10.1128/mcb.10.9.4872-4885.1990. Mol Cell Biol. 1990. PMID: 2201905 Free PMC article.
-
Identification of a DNA segment that is necessary and sufficient for alpha-specific gene control in Saccharomyces cerevisiae: implications for regulation of alpha-specific and a-specific genes.Mol Cell Biol. 1988 Jan;8(1):309-20. doi: 10.1128/mcb.8.1.309-320.1988. Mol Cell Biol. 1988. PMID: 3275872 Free PMC article.
-
Downstream activating sequence within the coding region of a yeast gene: specific binding in vitro of RAP1 protein.Mol Gen Genet. 1992 Dec;236(1):65-75. doi: 10.1007/BF00279644. Mol Gen Genet. 1992. PMID: 1494352
-
MAT alpha 1 can mediate gene activation by a-mating factor.Genes Dev. 1991 Oct;5(10):1924-34. doi: 10.1101/gad.5.10.1924. Genes Dev. 1991. PMID: 1916267
Cited by
-
Context-dependent function of the transcriptional regulator Rap1 in gene silencing and activation in Saccharomyces cerevisiae.Proc Natl Acad Sci U S A. 2023 Oct 3;120(40):e2304343120. doi: 10.1073/pnas.2304343120. Epub 2023 Sep 28. Proc Natl Acad Sci U S A. 2023. PMID: 37769255 Free PMC article.
-
Context dependent function of the transcriptional regulator Rap1 in gene silencing and activation in Saccharomyces cerevisiae.bioRxiv [Preprint]. 2023 May 11:2023.05.08.539937. doi: 10.1101/2023.05.08.539937. bioRxiv. 2023. Update in: Proc Natl Acad Sci U S A. 2023 Oct 3;120(40):e2304343120. doi: 10.1073/pnas.2304343120. PMID: 37214837 Free PMC article. Updated. Preprint.
-
Limits to transcriptional silencing in Saccharomyces cerevisiae.Genetics. 2023 Feb 9;223(2):iyac180. doi: 10.1093/genetics/iyac180. Genetics. 2023. PMID: 36495285 Free PMC article.
-
The Nuts and Bolts of Transcriptionally Silent Chromatin in Saccharomyces cerevisiae.Genetics. 2016 Aug;203(4):1563-99. doi: 10.1534/genetics.112.145243. Genetics. 2016. PMID: 27516616 Free PMC article. Review.
-
The nuclear pore complex mediates binding of the Mig1 repressor to target promoters.PLoS One. 2011;6(11):e27117. doi: 10.1371/journal.pone.0027117. Epub 2011 Nov 14. PLoS One. 2011. PMID: 22110603 Free PMC article.
References
Publication types
MeSH terms
Substances
Grants and funding
LinkOut - more resources
Full Text Sources
Molecular Biology Databases
Miscellaneous