CHARACTERIZATION OF TESTIS SPECIFIC HISTONE BINDING PROTEIN (NASP)

这项研究的目标是表征生物角色 睾丸和精子特异组蛋白结合蛋白NASP(核 精子发生过程中的自身抗原精子蛋白)。我们的假设是 组蛋白结合蛋白可以结合和运输 睾丸组蛋白从胞浆进入胞核期间 DNA复制完成后，减数分裂细胞分裂。通过 将睾丸组蛋白变体运送到细胞核中，如 组蛋白结合蛋白参与染色质和核小体 重组。NASP被认为是这样一种组蛋白结合 基于其睾丸特异性和核转位信号的蛋白质， 它与非洲爪哇组蛋白结合蛋白N1/N2的相似性，以及其 保守的组蛋白结合域。具体地说，我们问的是什么角色 NASP在睾丸特异的运动、运输和转移中的作用 组蛋白、从细胞质到细胞质的过渡蛋白或鱼精蛋白 细胞核和精母细胞或精子细胞的DNA。因此， 这项建议的具体目的是确定：1)具体的约束力 以及睾丸组蛋白、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/或体细胞组蛋白运送到哺乳动物体内 细胞核。3)NASP是否会将其结合的组蛋白转移到DNA上 与非洲爪哇组蛋白结合蛋白N1/N2的研究结果相同。4) 小鼠NASP的序列及其制备中的基因组结构 用于在小鼠身上进行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 包括睾丸组蛋白合成，过渡蛋白合成， 鱼精蛋白合成或所有这三种。

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.