Inoue A, Ishiji A, Kasagi S, Ishizuka M, Hirose S, Baba T, Hagiwara H
Research Center for Experimental Biology, Tokyo Institute of Technology, 4259 Nagatsuta-cho, Midori-ku, Yokohama, 226-8501, Japan.
Biochem Biophys Res Commun. 2000 Jul 5;273(2):398-403. doi: 10.1006/bbrc.2000.2953.
The cDNA for an RNA that is expressed predominantly in mouse spermatogenic cells was cloned and characterized. It was found to encode novel zinc finger protein. We first generated a cDNA fragment from mouse osteoblastic cells by the differential display method. To our surprise, Northern blot analysis revealed that the corresponding transcript was expressed at high levels in the testis rather than in osteoblastic cells. Therefore, using this fragment as a probe, we isolated the full-length cDNA (3340 bp) from a mouse testis cDNA library. Analysis of the open reading frame of the cDNA indicated that the encoded protein was a polypeptide of 942 amino acids residues that included three distinct domains, namely, a zinc finger domain of the Cys(2)-His(2) type, four basic amino acid-rich domains, and a myosin II-homology domain. In situ hybridization indicated that the transcript was present in seminiferous tubules of adult mice. Elevated expression of the transcript during testicular development in mice was restricted to spermatocytes at the pachytene stage of meiotic prophase and to round and elongated spermatids, as indicated by Northern blot analysis and RT-PCR. Our results suggest that this novel zinc finger protein might act as a transcriptional regulator during spermatogenesis and, in particular, during meiotic division.
克隆并鉴定了一种主要在小鼠生精细胞中表达的RNA的cDNA。发现它编码一种新型锌指蛋白。我们首先通过差异显示法从小鼠成骨细胞中生成了一个cDNA片段。令我们惊讶的是,Northern印迹分析显示相应的转录本在睾丸中高表达,而不是在成骨细胞中。因此,我们用这个片段作为探针,从小鼠睾丸cDNA文库中分离出了全长cDNA(3340 bp)。对该cDNA开放阅读框的分析表明,编码的蛋白质是一个由942个氨基酸残基组成的多肽,包含三个不同的结构域,即Cys(2)-His(2)型锌指结构域、四个富含碱性氨基酸的结构域和一个肌球蛋白II同源结构域。原位杂交表明该转录本存在于成年小鼠的生精小管中。Northern印迹分析和RT-PCR表明,在小鼠睾丸发育过程中,该转录本的表达升高仅限于减数分裂前期粗线期的精母细胞以及圆形和延长型精子细胞。我们的结果表明,这种新型锌指蛋白可能在精子发生过程中,特别是在减数分裂过程中作为转录调节因子发挥作用。