MOLECULAR CHARACTERIZATION OF TELOMERES IN HUMAN SPERM

描述：（改编自申请人的摘要）本项目的长期目标 建议是描述端粒的组织在 精子发生一个主要的重点是定义与 人类精子中的端粒染色体区域。的组件 端粒结合蛋白复合物（hsTBP）-与DS结合亲和力 六核苷酸重复序列（TTAGGG）端粒DNA -已被鉴定并提出 参与端粒与核膜的连接。一 hsTBP的组分之一，称为spH 2B，被认为是hsTBP的亚型。 体细胞组蛋白H2 B。hsRBP的另一个组分是一种新的50 kDa蛋白质 被称为P50。基于这些数据，申请人提出隔离， 克隆，并进一步表征spH 2B和p50。这种方法将 能够在人体内鉴定hsTBP的其他组分 生精细胞和精子。使用原子力显微镜的其他研究 和整体电子显微镜包括结构分析的 端粒DNA似乎保留其核小体样组织。的 后面的研究将探讨最近报道的T环的变化是否 用原位杂交技术观察了体细胞端粒的特征， 精子的交联端粒DNA制品。拟议的研究是 被认为对端粒酶活性的调节很重要， 减数分裂前期的同源染色体配对和重组， 受精后染色体被去除。

DESCRIPTION: (Adapted from the applicant's abstract) The long-term goal of this proposal is to characterize the organization of the telomere during spermatogenesis. A major emphasis is to define the molecules associated with the telomere chromosomal domains in human sperm. A component of the telomere-binding protein complex (hsTBP) - with binding affinity to the ds hexanucleotide repeat (TTAGGG) telomere DNA - has been identified and proposed to participate in the attachment of the telomeres to the nuclear envelope. One of the components of hsTBP, called spH2B, is regarded as a subtype of the somatic histone H2B. Another component of hsRBP is a novel 50 kDa protein identified as p50. Based on these data, the applicant proposes the isolation, cloning, and further characterization of spH2B and p50. This approach will enable the identification of additional components of hsTBP in human spermatogenic cells and sperm. Additional studies using atomic force microscopy and whole mount electron microscopy include a structural analysis of the telomeric DNA that appears to retain its nucleosomal-like organization. The latter studies will explore whether a variation of the recently reported t-loop characteristic of the telomere in somatic cells is observed using in situ cross-linked telomeric DNA preparations of sperm. The proposed studies are regarded as important for the regulation of telomerase activity during homologous chromosomal pairing and recombination during meiotic prophase and ordered chromosome withdrawal after fertilization.