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PMID: 9671481 Published · ppublish English Journal Article Research Support, Non-U.S. Gov't Research Support, U.S. Gov't, P.H.S.

Nuclear proteins Nut1p and Nut2p cooperate to negatively regulate a Swi4p-dependent lacZ reporter gene in Saccharomyces cerevisiae.

Molecular and cellular biology ·Vol. 18 ·No. 8 ·1998-08-00 ·Pages 4707-18

Tabtiang RK, Herskowitz I

Abstract

The URS2 region of the Saccharomyces cerevisiae HO upstream region contains 10 binding sites for the Swi4p/Swi6p transcription factor and confers Swi4p dependence for transcription. Using a hybrid promoter, UASGAL (upstream activation sequence of GAL1)-URS2R, in which the GAL1-10 regulatory region is fused to the proximal 360 bp of URS2, we isolated mutants in which Swi4p is no longer required for transcription. Mutations of SIN4, ROX3, SRB8, SRB9, SRB10, SRB11, and two novel genes, NUT1 and NUT2, relieve the requirement of Swi4p for expression of this reporter. We found that NUT1 (open reading frame [ORF] YGL151w) is a nonessential gene, that NUT2 (ORF YPR168w) is essential, and that both Nut1p and Nut2p encode nuclear proteins. Deletion of NUT1 causes a constitutive, Swi4p-independent phenotype only in combination with the nut2-1 allele or an allele of CCR4. In contrast, inactivation of a temperature-sensitive allele of NUT2, nut2-ts70, alone causes constitutivity. nut1Delta nut2-1 cells and sin4Delta cells exhibit Swi4p-independent expression of an ho-lacZ reporter but not of an intact ho gene. Likewise, a pPHO5-lacZ construct is constitutively expressed in nut1 nut2 mutants relative to their wild-type counterparts. These results suggest that Nut1p, Nut2p, Sin4p, and Ccr4p define a group of proteins that negatively regulate transcription in a subtle manner which is revealed by artificial reporter genes.

MeSH Terms
Alleles Amino Acid Sequence Animals Cell Nucleus/metabolism Cyclin-Dependent Kinase 8 Cyclin-Dependent Kinases/genetics Cyclins DNA-Binding Proteins Fungal Proteins/genetics,metabolism Gene Expression Regulation, Fungal Genes, Reporter Humans Lac Operon Mediator Complex Molecular Sequence Data Mutagenesis Nuclear Proteins/genetics,metabolism Phenotype RNA Polymerase II/genetics Ribonucleases Saccharomyces cerevisiae/genetics Saccharomyces cerevisiae Proteins Sequence Homology, Amino Acid Temperature Trans-Activators Transcription Factors/genetics,metabolism Transcription, Genetic
Chemicals
Cyclins DNA-Binding Proteins Fungal Proteins Mediator Complex NUT1 protein, S cerevisiae NUT2 protein, S cerevisiae Nuclear Proteins ROX3 protein, S cerevisiae SIN4 protein, S cerevisiae SSN2 protein, S cerevisiae SWI4 protein, S cerevisiae Saccharomyces cerevisiae Proteins Trans-Activators Transcription Factors SSN8 protein, S cerevisiae Cyclin-Dependent Kinase 8 Cyclin-Dependent Kinases SSN3 protein, S cerevisiae RNA Polymerase II CCR4 protein, S cerevisiae Ribonucleases
Authors & Affiliations
2 authors, click to expand affiliations / ORCID
Tabtiang R K
Program in Biochemistry and Molecular Biology, Department of Biochemistry and Biophysics, University of California, San Francisco, San Francisco, California 94143-0448, USA.
Herskowitz I
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Article Info
Journal
Molecular and cellular biology
Abbr.
Mol Cell Biol
ISSN
0270-7306
DOI
10.1128/MCB.18.8.4707
Published
1998-08-00
Pages
4707-18
Language
English
Region
United States
NLM ID
8109087
PMCID
PMC109057
Subset
IM
Grants
NIAID NIH HHS · AI18738 · United States
External Links
PubMed source DOI: 10.1128/MCB.18.8.4707 PMC Full Text View journal details
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