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PMID: 12370310 Published · ppublish English Journal Article

Identification of mZnf8, a mouse Krüppel-like transcriptional repressor, as a novel nuclear interaction partner of Smad1.

Molecular and cellular biology ·Vol. 22 ·No. 21 ·2002-11-00 ·Pages 7633-44

Jiao K, Zhou Y, Hogan BL

Abstract

To identify novel genes that play critical roles in mediating bone morphogenetic protein (BMP) signal pathways, we performed a yeast two-hybrid screen using Smad1 as bait. A novel mouse Krüppel-type zinc finger protein, mZnf8, was isolated. Interactions between mZnf8 and Smad proteins were further analyzed with various in vitro and in vivo approaches, including mammalian two-hybrid, in vitro glutathione S-transferase pulldown, and copurification assays. Results from functional analysis indicate that mZnf8 is a nuclear transcriptional repressor. Overexpression of mZnf8 represses activity of BMP and transforming growth factor beta (TGF-beta) reporters. Silencing the expression of endogenous mZnf8 with an RNA interference approach caused a significant increase in the expression of one BMP reporter. These results suggest that mZnf8 negatively regulates the TGF-beta/BMP signaling pathway in vivo. Transcription of mZnf8 is ubiquitous in mouse embryos, but high levels are specifically observed in adult mouse testes, with the same cell- and stage-specific transcription pattern as Smad1. Our data support the hypothesis that mZnf8 plays critical roles in mediating BMP signaling during spermatogenesis.

MeSH Terms
Amino Acid Sequence Animals Blotting, Northern Bone Morphogenetic Proteins/metabolism COS Cells Cell Nucleus/metabolism Cloning, Molecular DNA-Binding Proteins/metabolism Glutathione Transferase/metabolism Humans In Situ Hybridization Kruppel-Like Transcription Factors Luciferases/metabolism Male Mice Models, Biological Molecular Sequence Data Protein Binding RNA/metabolism Recombinant Fusion Proteins/metabolism Repressor Proteins/biosynthesis,chemistry,physiology Reverse Transcriptase Polymerase Chain Reaction Sequence Homology, Amino Acid Signal Transduction Smad Proteins Smad1 Protein Spermatogenesis Testis/embryology,metabolism Time Factors Trans-Activators/metabolism Transcription Factors/metabolism Transcription, Genetic Transfection Transforming Growth Factor beta/metabolism Two-Hybrid System Techniques
Chemicals
Bone Morphogenetic Proteins DNA-Binding Proteins Kruppel-Like Transcription Factors Recombinant Fusion Proteins Repressor Proteins SMAD1 protein, human Smad Proteins Smad1 Protein Smad1 protein, mouse Trans-Activators Transcription Factors Transforming Growth Factor beta ZNF8 protein, human Zfp128 protein, mouse RNA Luciferases Glutathione Transferase
Authors & Affiliations
3 authors, click to expand affiliations / ORCID
Jiao Kai
Howard Hughes Medical Institute and Department of Cell Biology, Vanderbilt University Medical School, Nashville, Tennessee 37232, USA.
Zhou Yingna
Hogan Brigid L M
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Article Info
Journal
Molecular and cellular biology
Abbr.
Mol Cell Biol
ISSN
0270-7306
DOI
10.1128/MCB.22.21.7633-7644.2002
Published
2002-11-00
Pages
7633-44
Language
English
Region
United States
NLM ID
8109087
PMCID
PMC135661
Subset
IM
External Links
PubMed source DOI: 10.1128/MCB.22.21.7633-7644.2002 PMC Full Text View journal details
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