Namekawa Satoshi, Hamada Fumika, Ishii Satomi, Ichijima Yosuke, Yamaguchi Taiki, Nara Takayuki, Kimura Seisuke, Ishizaki Takashi, Iwabata Kazuki, Koshiyama Akiyo, Teraoka Hirobumi, Sakaguchi Kengo
Department of Applied Biological Science, Faculty of Science and Technology, Tokyo University of Science, 2641 Yamazaki, Chiba 278-0022, Noda, Japan.
Biochim Biophys Acta. 2003 May 13;1627(1):47-55. doi: 10.1016/s0167-4781(03)00073-3.
DNA ligase I is thought to be essential for DNA replication, repair and recombination, at least in the mitotic cell cycle, but whether this is also the case during the meiotic cell cycle is still obscure. To investigate the role of DNA ligase I during the meiotic cell cycle, we cloned the Coprinus cinereus DNA ligase I cDNA (CcLIG1). Northern blotting analysis indicated that CcLIG1 is expressed not only in the premeiotic S-phase but also during the meiotic cell cycle itself. Especially, intense signals were observed in the leptotene and zygotene stages. Western blotting analysis indicated that CcLIG1 is expressed through the meiotic cell cycle and immunofluorescence also showed CcLIG1 protein staining in meiotic cells. Interestingly, the patterns was similar to that for the C. cinereus proliferating cell nuclear antigen gene (CcPCNA) and immunoprecipitation analysis suggested that CcPCNA binds to CcLIG1 in crude extracts of meiotic prophase I tissues. Based on these observations, relationships and roles during the meiotic cell cycle are discussed.
人们认为DNA连接酶I对于DNA复制、修复和重组至关重要,至少在有丝分裂细胞周期中是这样,但在减数分裂细胞周期中是否如此仍不清楚。为了研究DNA连接酶I在减数分裂细胞周期中的作用,我们克隆了灰盖鬼伞DNA连接酶I的cDNA(CcLIG1)。Northern印迹分析表明,CcLIG1不仅在减数分裂前的S期表达,而且在减数分裂细胞周期本身也有表达。特别是,在细线期和偶线期观察到强烈的信号。Western印迹分析表明,CcLIG1在整个减数分裂细胞周期中都有表达,免疫荧光也显示减数分裂细胞中有CcLIG1蛋白染色。有趣的是,这种模式与灰盖鬼伞增殖细胞核抗原基因(CcPCNA)的模式相似,免疫沉淀分析表明,在减数分裂前期I组织的粗提物中,CcPCNA与CcLIG1结合。基于这些观察结果,我们讨论了减数分裂细胞周期中的关系和作用。