CHARACTERIZATION OF TESTIS SPECIFIC HISTONE BINDING PROTEIN (NASP)

这项研究的目标是描述生物学作用的特征， 睾丸和精子特异性组蛋白结合蛋白NASP（核 自身抗原性精子蛋白）。 我们的假设是 一种组蛋白结合蛋白能够结合并运输 睾丸组织蛋白从细胞质进入细胞核的两个过程 DNA复制完成后的减数分裂。 通过 将睾丸组蛋白变体转运到细胞核中，例如 组蛋白结合蛋白参与染色质和核小体 重组。 假设NASP是这样的组蛋白结合蛋白， 蛋白质基于其睾丸特异性其核转位信号， 它与非洲爪蟾组蛋白结合蛋白N1/N2相似， 保守的组蛋白结合域。 具体来说，我们要问的是 NASP参与睾丸特异性细胞的运动、转运和转移 组蛋白、过渡蛋白或鱼精蛋白从细胞质转移到 细胞核和精母细胞或精子细胞的DNA。 因此 本提案的具体目的是确定：（1）具体约束力 睾丸组蛋白、过渡蛋白和鱼精蛋白的亲和力 以及NASP的体细胞组蛋白。 2)NASP是否真的 将结合的睾丸和/或体细胞组蛋白转运到哺乳动物中 细胞核 3)NASP是否会将其结合的组蛋白转移到DNA中 这已经在爪蟾组蛋白结合蛋白N1/N2中得到证实。 四、 小鼠NASP的序列测定及其基因组结构分析 用于小鼠的阶段特异性和基因靶向研究。 5)是否 小鼠精子发生过程中NASP的合成是协调的 睾丸组蛋白合成，过渡蛋白合成， 鱼精蛋白合成或这三种。

The goal of this research is the characterization of the biological role of the testis and sperm specific histone binding protein NASP (nuclear autoantigenic sperm protein) during spermatogenesis. Our hypothesis is that a histone-binding protein could function to bind and transport testicular histone from the cytoplasm into the nucleus during the two meiotic cell divisions after DNA replication has finished. By transporting testicular histone variants into the nucleus, such as histone-binding protein would participate in chromatin and nucelosome reorganization. NASP is hypothesized to be such a histone-binding protein based on its testis specificity its nuclear translocation signal, its similarity to the Xenopus histone binding protein N1/N2, and its conserved histone binding domains. Specifically, we are asking what role NASP plays in the movement, transport and transfer of testis specific histones, transition proteins or protamines from the cytoplasm to the nucleus and to the DNA of the spermatocyte or spermatid. Therefore, the Specific Aims of this proposal are to determine: 1) The specific binding and affinity of testicular histones, transition proteins, and protamines as well as somatic histones for NASP. 2) Whether NASP actually transports bound testicular and/or somatic histones into the mammalian cell nucleus. 3) Whether NASP will transfer its bound histones to DNA as has been demonstrated for Xenopus histone binding protein N1/N2. 4) The sequence of mouse NASP and its genomic organization in preparation for stage specific and gene targeting studies in the mouse. 5) Whether the synthesis of NASP during spermatogenesis in the mouse is coordinated with testicular histone synthesis, transition protein synthesis, protamine synthesis or all three of these.