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Molecular and Cellular Biology, November 2002, p. 7633-7644, Vol. 22, No. 21
0270-7306/02/$04.00+0     DOI: 10.1128/MCB.22.21.7633-7644.2002
Copyright © 2002, American Society for Microbiology. All Rights Reserved.

Identification of mZnf8, a Mouse Krüppel-Like Transcriptional Repressor, as a Novel Nuclear Interaction Partner of Smad1

Kai Jiao, Yingna Zhou, and Brigid L. M. Hogan*

Howard Hughes Medical Institute and Department of Cell Biology, Vanderbilt University Medical School, Nashville, Tennessee 37232

Received 27 March 2002/ Returned for modification 22 May 2002/ Accepted 8 August 2002

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-ß) 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-ß/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.

* Corresponding author. Mailing address: Vanderbilt University School of Medicine, C-2310 Medical Center North, Nashville, TN 37232. Phone: (615) 343-6418. Fax: (615) 343-2033. E-mail: Brigid.Hogan{at}mcmail.vanderbilt.edu.

Molecular and Cellular Biology, November 2002, p. 7633-7644, Vol. 22, No. 21
0022-538X/02/$04.00+0     DOI: 10.1128/MCB.22.21.7633-7644.2002
Copyright © 2002, American Society for Microbiology. All Rights Reserved.



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