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Mol Cell Biol, April 1998, p. 2118-2129, Vol. 18, No. 4
0270-7306/98/$04.00+0
Copyright © 1998, American Society for Microbiology. All rights reserved.

The Schizosaccharomyces pombe mei4+ Gene Encodes a Meiosis-Specific Transcription Factor Containing a forkhead DNA-Binding Domain

S. Horie,1,dagger Y. Watanabe,2,Dagger K. Tanaka,1,§ S. Nishiwaki,1,par H. Fujioka,1,# H. Abe,1 M. Yamamoto,2 and C. Shimoda1,*

Department of Biology, Faculty of Science, Osaka City University, Sumiyoshi-ku, Osaka 558,1 and Department of Biophysics and Biochemistry, Graduate School of Science, University of Tokyo, Bunkyo-ku, Tokyo 113,2 Japan

Received 29 October 1997/Returned for modification 4 December 1997/Accepted 22 December 1997

The mei4+ gene of the fission yeast Schizosaccharomyces pombe was cloned by functional complementation. The mei4 disruptant failed to complete meiosis-I but could proliferate normally. mei4+ was transcribed only in meiosis-proficient diploid cells after premeiotic DNA replication. The mei4+ open reading frame encodes a 57-kDa serine-rich protein comprised of 517 amino acids with a forkhead/HNF3 DNA-binding domain in the amino-terminal region. Transcription of spo6+, a gene required for sporulation, was dependent on the mei4+ function. Two copies of the GTAAAYA consensus sequence, proposed as the binding site for human forkhead proteins, were found in the promoter region of spo6+. A gel mobility shift assay demonstrated the sequence-dependent binding of the GST-Mei4 forkhead domain fusion protein to DNA fragments with one of the consensus elements. Deletion of this consensus element from the spo6 promoter abolished the transcription of spo6+ and resulted in a sporulation deficiency. One-hybrid assay of Mei4 which was fused to the Gal4 DNA-binding domain localized the transcriptional activation domain in the C-terminal 140 amino acids of Mei4. These results indicate that Mei4 functions as a meiosis-specific transcription factor of S. pombe.

* Corresponding author. Mailing address: Department of Biology, Faculty of Science, Osaka City University, Sumiyoshi-ku, Osaka 558, Japan. Phone: 81 6605 2576. Fax: 81 6605 3158. E-mail: shimoda{at}sci.osaka-cu.ac.jp.

dagger Present address: Biotechnology Research laboratories, Takara Shuzo Co., Ltd., Otsu, Shiga 520-21, Japan.

Dagger Present address: Cell Cycle Laboratory, Imperial Cancer Research Fund, London WC2A 3PX, United Kingdom.

§ Present address: Department of Biochemistry, Faculty of Medicine, University of Tokyo, Bunkyo-ku, Tokyo 113, Tokyo, Japan.

par Present address: Chemical Products Research Laboratory, Fujisawa Pharmaceutical Co., Ltd., Tsukuba, Ibaraki 300-26, Japan.

# Present address: Asahi Breweries Co., Nishinomiya, Hyogo 663, Japan.




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