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PMID: 15684403 Published · ppublish English Journal Article Research Support, Non-U.S. Gov't Research Support, U.S. Gov't, P.H.S.

Role for SUMO modification in facilitating transcriptional repression by BKLF.

Molecular and cellular biology ·Vol. 25 ·No. 4 ·2005-02-00 ·Pages 1549-59

Perdomo J, Verger A, Turner J, Crossley M

Abstract

Small ubiquitin-like modifier (SUMO) is a protein moiety that is ligated to lysine residues on a variety of target proteins. Many known SUMO substrates are transcription factors or coregulators of transcription, and in most cases, modification with SUMO leads to the attenuation of transcriptional activation. We have examined basic Kruppel-like factor/Kruppel-like factor 3 (BKLF), a zinc finger transcription factor that is known to function as a potent transcriptional repressor. We show that BKLF recruits the E2 SUMO-conjugating enzyme Ubc9 and can be modified by the addition of SUMO-1 in vitro and in vivo. The SUMO E3 ligases PIAS1, PIASgamma, PIASxalpha, and PIASxbeta but not Pc2 enhance the sumoylation of BKLF. Site-directed mutagenesis identified two lysines (K10 and K197) of BKLF as the sumoylation sites. Sumoylation does not detectably affect DNA binding by BKLF, but mutation of the sumoylation sites reduces transcriptional repression activity. Most interestingly, when mutations preventing sumoylation are combined with an additional mutation that eliminates contact with the C-terminal binding protein (CtBP) corepressor, BKLF becomes an activator of transcription. These results link SUMO modification to transcriptional repression and demonstrate that both recruitment of CtBP and sumoylation are required for full repression by BKLF.

MeSH Terms
Alcohol Oxidoreductases Animals COS Cells Cells, Cultured Chlorocebus aethiops DNA-Binding Proteins/genetics,metabolism Drosophila melanogaster/metabolism Lysine/metabolism Mice Mutagenesis, Site-Directed/genetics Phosphoproteins/metabolism Protein Inhibitors of Activated STAT Proteins/genetics,metabolism SUMO-1 Protein/genetics,metabolism Transcription, Genetic/genetics Two-Hybrid System Techniques
Chemicals
DNA-Binding Proteins Phosphoproteins Pias1 protein, mouse Protein Inhibitors of Activated STAT Proteins SUMO-1 Protein Alcohol Oxidoreductases C-terminal binding protein Lysine
Authors & Affiliations
4 authors, click to expand affiliations / ORCID
Perdomo José
School of Molecular and Microbial Biosciences G08, University of Sydney, Sydney, New South Wales 2006, Australia.
Verger Alexis
Turner Jeremy
Crossley Merlin
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Article Info
Journal
Molecular and cellular biology
Abbr.
Mol Cell Biol
ISSN
0270-7306
DOI
10.1128/MCB.25.4.1549-1559.2005
Published
2005-02-00
Pages
1549-59
Language
English
Region
United States
NLM ID
8109087
PMCID
PMC548027
Subset
IM
External Links
PubMed source DOI: 10.1128/MCB.25.4.1549-1559.2005 PMC Full Text View journal details
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