Identification of a novel Leucine-rich repeat protein and candidate PP1 regulatory subunit expressed in developing spermatids.
Identification of a novel Leucine-rich repeat protein and candidate PP1 regulatory subunit expressed in developing spermatids.
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DOI：
10.1186/1471-2121-9-9
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发表时间：
2008-01-31
期刊：
影响因子：
--
通讯作者：
Sperry AO
中科院分区：
文献类型：
--
作者：
Wang R;Sperry AO
精子发生是由一系列高度调控的发育变化组成的，这些变化将前体生殖细胞转化为高度特化的精子。精子发生的最后阶段，称为精子发生，涉及巨大的形态变化，包括顶体的形成，细胞核的伸长和凝聚，鞭毛的形成，以及不必要的细胞质的处理。发育中的精子细胞骨架的一个重要组成部分是manchette，这是一种独特的微管结构，围绕着发育中的精子细胞的细胞核，被认为有助于细胞核的重塑和精子细胞质的重新分配。尽管分子马达KIFC1已被证明与manchette相关，但其在manchette功能中的确切作用及其睾丸特异性蛋白伴侣的身份尚不清楚。本研究的目的是利用酵母2杂交筛选睾丸cDNA文库，鉴定睾丸中与KIFC1相互作用的蛋白质。在我们的筛选中发现的30%的相互作用克隆含有相同的编码40 kD蛋白的cDNA。这种相互作用蛋白在其氨基末端有4个富含亮氨酸的重复序列，主要在睾丸中表达；因此，我们将这种蛋白命名为睾丸富含亮氨酸重复蛋白或TLRR。通过亲和层析还发现TLRR与KIFC1靶向结构域紧密结合。除了富含亮氨酸的重复序列外，TLRR还含有蛋白磷酸酶-1 （PP1）的一致结合位点。使用TLRR特异性抗体的免疫细胞化学表明，该蛋白在发育中的精子的manchette附近被发现。我们已经发现了一种以前未被发现的富含亮氨酸的重复蛋白，该蛋白在睾丸中大量表达，并可能通过与KIFC1分子马达的相互作用与发育中的精子的manchette相关。TLRR与PP1的一类调控亚基同源，PP1是蛋白质可逆磷酸化的中心磷酸酶，是调节许多细胞内过程的关键。TLRR可能针对发育中的精子细胞核附近的这一重要信号分子，以控制精子发生过程中的细胞重排。
Spermatogenesis is comprised of a series of highly regulated developmental changes that transform the precursor germ cell into a highly specialized spermatozoon. The last phase of spermatogenesis, termed spermiogenesis, involves dramatic morphological change including formation of the acrosome, elongation and condensation of the nucleus, formation of the flagella, and disposal of unnecessary cytoplasm. A prominent cytoskeletal component of the developing spermatid is the manchette, a unique microtubular structure that surrounds the nucleus of the developing spermatid and is thought to assist in both the reshaping of the nucleus and redistribution of spermatid cytoplasm. Although the molecular motor KIFC1 has been shown to associate with the manchette, its precise role in function of the manchette and the identity of its testis specific protein partners are unknown. The purpose of this study was to identify proteins in the testis that interact with KIFC1 using a yeast 2 hybrid screen of a testis cDNA library. Thirty percent of the interacting clones identified in our screen contain an identical cDNA encoding a 40 kD protein. This interacting protein has 4 leucine-rich repeats in its amino terminal half and is expressed primarily in the testis; therefore we have named this protein testis leucine-rich repeat protein or TLRR. TLRR was also found to associate tightly with the KIFC1 targeting domain using affinity chromatography. In addition to the leucine-rich repeats, TLRR contains a consensus-binding site for protein phosphatase-1 (PP1). Immunocytochemistry using a TLRR specific antibody demonstrates that this protein is found near the manchette of developing spermatids. We have identified a previously uncharacterized leucine-rich repeat protein that is expressed abundantly in the testis and associates with the manchette of developing spermatids, possibly through its interaction with the KIFC1 molecular motor. TLRR is homologous to a class of regulatory subunits for PP1, a central phosphatase in the reversible phosphorylation of proteins that is key to modulation of many intracellular processes. TLRR may serve to target this important signaling molecule near the nucleus of developing spermatids in order to control the cellular rearrangements of spermiogenesis.
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影响因子：
14.9
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