Zhu X, Naz R K
Department of Obstetrics and Gynecology, Medical College of Ohio, Richard D. Ruppert Health Center, Toledo 43614, USA.
Biochem Biophys Res Commun. 1998 Aug 10;249(1):56-60. doi: 10.1006/bbrc.1998.9052.
A new isoform of cyclin I, designated cyclin ITI, was cloned from the human testis cDNA-lambda gt10 library by using a approximately 0.5 kb PCR fragment that was obtained by using primers based on the published cyclin I sequence from the human brain. The cyclin ITI cDNA is 1443-bp long and has an ORF of 178 aa with the first ATG Met start codon at nt 1 and has the well-conserved sequence of cyclin box. The 5' flanking sequence of 728-bp has a high similarity (99.6%) with the 5'-intron of the human beta-polymerase gene. The nt sequence of cyclin ITI cDNA has high similarity (99.3%) with the cyclin I gene sequence. The cyclin ITI was found to have three amino acids mutations at nt 25, 172, 223, respectively, including one in the well-conserved sequence of cyclin box. Northern blot analysis indicated that the cyclin ITI is expressed in human testis at increased level compared to other tissues. The in vitro translated protein of the cyclin ITI cDNA was recognized specifically by the antibodies raised against the human sperm proteins, indicating its presence in human sperm. This new isoform may have a physiological role in spermatogenesis and/or human sperm cell function--especially in capacitation and/or acrosome reaction.
通过使用基于已发表的人脑细胞周期蛋白I序列设计的引物获得的约0.5 kb PCR片段,从人睾丸cDNA - λgt10文库中克隆出一种新的细胞周期蛋白I亚型,命名为细胞周期蛋白ITI。细胞周期蛋白ITI cDNA长1443 bp,有一个178个氨基酸的开放阅读框,起始密码子ATG Met位于第1个核苷酸处,并且具有细胞周期蛋白框的保守序列。728 bp的5'侧翼序列与人β - 聚合酶基因的5'内含子具有高度相似性(99.6%)。细胞周期蛋白ITI cDNA的核苷酸序列与细胞周期蛋白I基因序列具有高度相似性(99.3%)。发现细胞周期蛋白ITI在核苷酸25、172、223处分别有三个氨基酸突变,其中一个在细胞周期蛋白框的保守序列中。Northern印迹分析表明,与其他组织相比,细胞周期蛋白ITI在人睾丸中表达水平升高。细胞周期蛋白ITI cDNA的体外翻译蛋白能被针对人精子蛋白产生的抗体特异性识别,表明其存在于人精子中。这种新亚型可能在精子发生和/或人精子细胞功能中具有生理作用——特别是在获能和/或顶体反应中。