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PMID: 12923057 Published · ppublish English Journal Article Research Support, Non-U.S. Gov't Research Support, U.S. Gov't, P.H.S.

Association of the RENT complex with nontranscribed and coding regions of rDNA and a regional requirement for the replication fork block protein Fob1 in rDNA silencing.

Genes & development ·Vol. 17 ·No. 17 ·2003-09-01 ·Pages 2162-76

Huang J, Moazed D

Abstract

Silencing within the yeast rDNA repeats inhibits hyperrecombination, represses transcription from foreign promoters, and extends replicative life span. rDNA silencing is mediated by a Sir2-containing complex called RENT (regulator of nucleolar silencing and telophase exit). We show that the Net1 (also called Cfi1) and Sir2 subunits of RENT localize primarily to two distinct regions within rDNA: in one of the nontranscribed spacers (NTS1) and around the Pol I promoter, extending into the 35S rRNA coding region. Binding to NTS1 overlaps the recombination hotspot and replication fork barrier elements, which have been shown previously to require the Fob1 protein for their activities. In cells lacking Fob1, silencing and the association of RENT subunits are abolished specifically at NTS1, while silencing and association at the Pol I promoter region are unaffected or increased. We find that Net1 and Sir2 are physically associated with Fob1 and subunits of RNA polymerase I. Together with the localization data, these results suggest the existence of two distinct modes for the recruitment of the RENT complex to rDNA and reveal a role for Fob1 in rDNA silencing and in the recruitment of the RENT complex. Furthermore, the Fob1-dependent associations of Net1 and Sir2 with the recombination hotspot region strongly suggest that Sir2 acts directly at this region to carry out its inhibitory effect on rDNA recombination and accelerated aging.

MeSH Terms
Cell Cycle Proteins/genetics,metabolism DNA, Ribosomal DNA-Binding Proteins/metabolism Gene Silencing/physiology Histone Deacetylases/genetics,metabolism Nuclear Proteins/genetics,metabolism RNA Polymerase I/metabolism Saccharomyces cerevisiae/genetics,metabolism Saccharomyces cerevisiae Proteins/genetics,metabolism Silent Information Regulator Proteins, Saccharomyces cerevisiae/genetics,metabolism Sirtuin 2 Sirtuins/genetics,metabolism
Chemicals
Cell Cycle Proteins DNA, Ribosomal DNA-Binding Proteins FOB1 protein, S cerevisiae Net1 protein, S cerevisiae Nuclear Proteins Saccharomyces cerevisiae Proteins Silent Information Regulator Proteins, Saccharomyces cerevisiae RNA Polymerase I SIR2 protein, S cerevisiae Sirtuin 2 Sirtuins Histone Deacetylases
Authors & Affiliations
2 authors, click to expand affiliations / ORCID
Huang Julie
Department of Cell Biology, Harvard Medical School, Boston, MA 02115, USA.
Moazed Danesh
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Article Info
Journal
Genes & development
Abbr.
Genes Dev
ISSN
0890-9369
DOI
10.1101/gad.1108403
Published
2003-09-01
Epub
2003-00-15
Pages
2162-76
Language
English
Region
United States
NLM ID
8711660
PMCID
PMC196457
Subset
IM
External Links
PubMed source DOI: 10.1101/gad.1108403 PMC Full Text View journal details
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