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Rosbin: A Novel Homeobox-Like Protein Gene Expressed Exclusively in Round Spermatids

Department of Science for Laboratory Animal Experimentation,² Research Institute for Microbial Diseases, Osaka University, Osaka 565-0871, Japan Department of Urology,³ Osaka University Graduate School of Medicine, Osaka University, Osaka 565-0871, Japan Department of Anatomy and Developmental Biology,⁴ Graduate School of Medicine, Chiba University,Chiba 260-8670, Japan

Mammalian spermiogenesis is a complex process occurring in a highly coordinated fashion within the seminiferous tubules. To elucidate the molecular mechanisms controlling haploid germ cell differentiation, we have isolated haploid germ cell- specific cDNA clones from a subtracted cDNA library of mouse testis. One of these cDNAs, Rosbin, is 3.2 kilobases (kb) long and has an open reading frame of 2385 nucleotides encoding a putative protein of 795 amino acid residues. A computer-mediated homology search revealed that it contained a domain similar to that of homeobox genes. Northern blot analysis revealed a 3.2-kb mRNA expressed exclusively in male germ cells. Transcription of the Rosbin gene was not observed in prepubertal testis but became detectable after Day 23. By Western blot analysis the protein encoded by this gene had a molecular mass of 89 kDa, expressing specifically in the testis and localized to the nucleus of stages IV–VIII haploid round spermatids, predominantly at stages VII–VIII of spermatogenesis. ROSBIN is associated with and is most likely phosphorylated by protein kinase A. We suggest that it plays an important role in transcriptional regulation in haploid germ cells.

¹ Correspondence: Yoshitake Nishimune, Department of Science for Laboratory Animal Experimentation, Research Institute for Microbial Diseases, Osaka University, 3-1 Yamadaoka, Suita, Osaka 565-0871, Japan. FAX: 81 6 6879 8339, nishimun{at}biken.osaka-u.ac.jp

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