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PMID: 7744964 Published · ppublish English Journal Article Research Support, Non-U.S. Gov't Research Support, U.S. Gov't, Non-P.H.S. Research Support, U.S. Gov't, P.H.S.

The carboxy termini of Sir4 and Rap1 affect Sir3 localization: evidence for a multicomponent complex required for yeast telomeric silencing.

The Journal of cell biology ·Vol. 129 ·No. 4 ·1995-05-00 ·Pages 909-24

Cockell M, Palladino F, Laroche T, Kyrion G, Liu C, Lustig AJ, Gasser SM

Abstract

The Silent Information Regulatory proteins, Sir3 and Sir4, and the telomeric repeat-binding protein RAP1 are required for the chromatin-mediated gene repression observed at yeast telomeric regions. All three proteins are localized by immunofluorescence staining to foci near the nuclear periphery suggesting a relationship between subnuclear localization and silencing. We present several lines of immunological and biochemical evidence that Sir3, Sir4, and RAP1 interact in intact yeast cells. First, immunolocalization of Sir3 to foci at the yeast nuclear periphery is lost in rap1 mutants carrying deletions for either the terminal 28 or 165 amino acids of RAP1. Second, the perinuclear localization of both Sir3 and RAP1 is disrupted by overproduction of the COOH terminus of Sir4. Third, overproduction of the Sir4 COOH terminus alters the solubility properties of both Sir3 and full-length Sir4. Finally, we demonstrate that RAP1 and Sir4 coprecipitate in immune complexes using either anti-RAP1 or anti-Sir4 antibodies. We propose that the integrity of a tertiary complex between Sir4, Sir3, and RAP1 is involved in both the maintenance of telomeric repression and the clustering of telomeres in foci near the nuclear periphery.

MeSH Terms
Cell Compartmentation Cell Nucleus/metabolism Fluorescent Antibody Technique Fungal Proteins/genetics,metabolism Macromolecular Substances Nuclear Proteins/genetics,metabolism Peptide Fragments/genetics,metabolism Precipitin Tests Protein Binding Recombinant Proteins/metabolism Saccharomyces cerevisiae/cytology,genetics Silent Information Regulator Proteins, Saccharomyces cerevisiae Solubility Structure-Activity Relationship Telomere/genetics Trans-Activators/genetics,metabolism Transcription Factors/isolation & purification,metabolism
Chemicals
Fungal Proteins Macromolecular Substances Nuclear Proteins Peptide Fragments Recombinant Proteins SIR3 protein, S cerevisiae SIR4 protein, S cerevisiae Silent Information Regulator Proteins, Saccharomyces cerevisiae Trans-Activators Transcription Factors
Authors & Affiliations
7 authors, click to expand affiliations / ORCID
Cockell M
Swiss Institute for Experimental Cancer Research (ISREC), Lausanne.
Palladino F
Laroche T
Kyrion G
Liu C
Lustig A J
Gasser S M
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Article Info
Journal
The Journal of cell biology
Abbr.
J Cell Biol
ISSN
0021-9525
DOI
10.1083/jcb.129.4.909
Published
1995-05-00
Pages
909-24
Language
English
Region
United States
NLM ID
0375356
PMCID
PMC2120499
Subset
IM
Grants
NCI NIH HHS · P30CA08748 · United States
External Links
PubMed source DOI: 10.1083/jcb.129.4.909 PMC Full Text View journal details
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