Faculty Collaboration Database

Sequence-specific transcriptional repression by KS1, a multiple-zinc-finger-Krüppel-associated box protein. Mol Cell Biol 2001 Feb;21(3):928-39

Date

01/12/2001

Pubmed ID

11154279

Pubmed Central ID

PMC86683

DOI

10.1128/MCB.21.3.928-939.2001

Scopus ID

2-s2.0-0035136902 (requires institutional sign-in at Scopus site)   67 Citations

Abstract

The vertebrate genome contains a large number of Krüppel-associated box-zinc finger genes that encode 10 or more C(2)-H(2) zinc finger motifs. Members of this gene family have been proposed to function as transcription factors by binding DNA through their zinc finger region and repressing gene expression via the KRAB domain. To date, however, no Krüppel-associated box-zinc finger protein (KRAB-ZFP) and few proteins with 10 or more zinc finger motifs have been shown to bind DNA in a sequence-specific manner. Our laboratory has recently identified KS1, a member of the KRAB-ZFP family that contains 10 different C(2)-H(2) zinc finger motifs, 9 clustered at the C terminus with an additional zinc finger separated by a short linker region. In this study, we used a random oligonucleotide binding assay to identify a 27-bp KS1 binding element (KBE). Reporter assays demonstrate that KS1 represses the expression of promoters containing this DNA sequence. Deletion and site-directed mutagenesis reveal that KS1 requires nine C-terminal zinc fingers and the KRAB domain for transcriptional repression through the KBE site, whereas the isolated zinc finger and linker region are dispensable for this function. Additional biochemical assays demonstrate that the KS1 KRAB domain interacts with the KAP-1 corepressor, and mutations that abolish this interaction alleviate KS1-mediated transcriptional repression. Thus, this study provides the first direct evidence that a KRAB-ZFP binds DNA to regulate gene expression and provides insight into the mechanisms used by multiple-zinc-finger proteins to recognize DNA sequences.

Author List

Gebelein B, Urrutia R

Author

Raul A. Urrutia MD Center Director, Professor in the Surgery department at Medical College of Wisconsin

MESH terms used to index this publication - Major topics in bold

Animals
Base Sequence
Binding Sites
CHO Cells
Cricetinae
DNA
DNA Primers
DNA-Binding Proteins
Humans
Promoter Regions, Genetic
Recombinant Fusion Proteins
Repressor Proteins
Transcription Factors
Transfection
Tripartite Motif-Containing Protein 28
Zinc Fingers