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PMID: 11432837 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.

Spt16-Pob3 and the HMG protein Nhp6 combine to form the nucleosome-binding factor SPN.

The EMBO journal ·Vol. 20 ·No. 13 ·2001-07-02 ·Pages 3506-17

Formosa T, Eriksson P, Wittmeyer J, Ginn J, Yu Y, Stillman DJ

Abstract

Yeast Spt16/Cdc68 and Pob3 form a heterodimer that acts in both DNA replication and transcription. This is supported by studies of new alleles of SPT16 described here. We show that Spt16-Pob3 enhances HO transcription through a mechanism that is affected by chromatin modification, since some of the defects caused by mutations can be suppressed by deleting the histone deacetylase Rpd3. While otherwise conserved among many eukaryotes, Pob3 lacks the HMG1 DNA-binding motif found in similar proteins such as the SSRP1 subunit of human FACT. SPT16 and POB3 display strong genetic interactions with NHP6A/B, which encodes an HMG1 motif, suggesting that these gene products function coordinately in vivo. While Spt16-Pob3 and Nhp6 do not appear to form stable heterotrimers, Nhp6 binds to nucleosomes and these Nhp6-nucleosomes can recruit Spt16-Pob3 to form SPN-nucleosomes. These complexes have altered electrophoretic mobility and a distinct pattern of enhanced sensitivity to DNase I. These results suggest that Spt16-Pob3 and Nhp6 cooperate to function as a novel nucleosome reorganizing factor.

MeSH Terms
Alleles Amino Acid Substitution Antigens, Nuclear Carrier Proteins/genetics,metabolism Cell Cycle Proteins/genetics,metabolism DNA-Binding Proteins/genetics,metabolism Fungal Proteins/genetics,metabolism Gene Expression Regulation, Fungal Genotype HMGN Proteins High Mobility Group Proteins/genetics,metabolism Histones/metabolism Humans Mutagenesis, Site-Directed Nuclear Matrix-Associated Proteins Nuclear Proteins/genetics,metabolism Nucleosomes/genetics,physiology Phenotype Protein Subunits Recombinant Proteins/metabolism Saccharomyces cerevisiae/genetics,physiology Saccharomyces cerevisiae Proteins Spindle Apparatus Transcription Factors Transcription, Genetic Transcriptional Elongation Factors
Chemicals
Antigens, Nuclear Carrier Proteins Cell Cycle Proteins DNA-Binding Proteins Fungal Proteins HMGN Proteins High Mobility Group Proteins Histones NHP6A protein, S cerevisiae NHP6B protein, S cerevisiae NUMA1 protein, human Nuclear Matrix-Associated Proteins Nuclear Proteins Nucleosomes POB3 protein, S cerevisiae Protein Subunits Recombinant Proteins SPT16 protein, S cerevisiae SSRP1 protein, human SUPT16H protein, human Saccharomyces cerevisiae Proteins Transcription Factors Transcriptional Elongation Factors
Authors & Affiliations
6 authors, click to expand affiliations / ORCID
Formosa T
Department of Biochemistry, University of Utah School of Medicine, 50 N. Medical Drive Rm 211, Salt Lake City, UT 84132, USA. [email protected]
Eriksson P
Wittmeyer J
Ginn J
Yu Y
Stillman D J
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Article Info
Journal
The EMBO journal
Abbr.
EMBO J
ISSN
0261-4189
DOI
10.1093/emboj/20.13.3506
Email
[email protected]
Published
2001-07-02
Pages
3506-17
Language
English
Region
England
NLM ID
8208664
PMCID
PMC125512
Subset
IM
External Links
PubMed source DOI: 10.1093/emboj/20.13.3506 PMC Full Text View journal details
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