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一种编码在睾丸单倍体生殖细胞中特异性表达的DNA结合蛋白(Hils1)的新型cDNA的分离与鉴定。

Isolation and characterization of a novel cDNA encoding a DNA-binding protein (Hils1) specifically expressed in testicular haploid germ cells.

作者信息

Iguchi Naoko, Tanaka Hiromitsu, Yomogida Kentaro, Nishimune Yoshitake

机构信息

Department of Science for Laboratory Animal Experimentation, Research Institute for Microbial Diseases, Osaka University, Suita City, Osaka, Japan.

出版信息

Int J Androl. 2003 Dec;26(6):354-65. doi: 10.1046/j.0105-6263.2003.00449.x.

Abstract

A cDNA encoding a protein homologous with histone H1 has been cloned from a haploid germ cell specific cDNA library. Deduced amino acid sequence (170 amino acids) showed 40% identity with histone H1 globular domain. Messenger RNA of the gene was observed exclusively in the testis, and was accumulated after post-natal day 23. Western blotting analysis showed that the protein encoded by this gene is about 19 kDa in molecular weight, and it was exclusively recovered from the nuclei of testicular germ cells. Immunohistochemical analysis showed that the protein was localized to the nuclei of round and elongating spermatids, consistent with the results of immunoblot analysis. Thus, the gene product was named Hils1 (histone H1 like protein in spermatids 1). In vitro DNA-binding experiments using DNA-cellulose mini-columns showed that Hils1 was able to bind to both double and single stranded-DNAs in a non-sequence-specific manner. These findings suggest that Hils1 may play an important role in the structural changes of spermatid nuclei, such as nuclear condensation, and gene regulation of haploid germ cell differentiation.

摘要

已从单倍体生殖细胞特异性cDNA文库中克隆出一个编码与组蛋白H1同源蛋白的cDNA。推导的氨基酸序列(170个氨基酸)与组蛋白H1球状结构域有40%的同一性。该基因的信使RNA仅在睾丸中观察到,并在出生后第23天开始积累。蛋白质印迹分析表明,该基因编码的蛋白质分子量约为19 kDa,且仅从睾丸生殖细胞核中回收。免疫组织化学分析表明,该蛋白质定位于圆形和伸长的精子细胞核,与免疫印迹分析结果一致。因此,该基因产物被命名为Hils1(精子细胞中的组蛋白H1样蛋白1)。使用DNA-纤维素微型柱进行的体外DNA结合实验表明,Hils1能够以非序列特异性方式与双链和单链DNA结合。这些发现表明,Hils1可能在精子细胞核的结构变化(如核浓缩)以及单倍体生殖细胞分化的基因调控中发挥重要作用。

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