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Jiao, K, Zhou, Y and Hogan, BL (2002) Identification of mZnf8, a mouse Krüppel-like transcriptional repressor, as a novel nuclear interaction partner of Smad1. Mol. Cell. Biol. 22:7633-44
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.
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; Repressor Proteins/chemistry; Repressor Proteins/physiology; Reverse Transcriptase Polymerase Chain Reaction; Sequence Homology, Amino Acid; Signal Transduction; Smad Proteins; Smad1 Protein; Spermatogenesis; Testis/embryology; Testis/metabolism; Time Factors; Trans-Activators/metabolism; Transcription Factors/metabolism; Transcription, Genetic; Transfection; Transforming Growth Factor beta/metabolism; Two-Hybrid System Techniques
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