Okuda Hidenobu, DeBoer Kathleen, O'Connor Anne E, Merriner D Jo, Jamsai Duangporn, O'Bryan Moira K
Monash Biomedicine Discovery Institute, Monash University, Clayton, Victoria, Australia; and.
Department of Anatomy and Developmental Biology, Monash University, Clayton, Victoria, Australia.
FASEB J. 2017 Mar;31(3):1141-1152. doi: 10.1096/fj.201600909R. Epub 2016 Dec 21.
Infertility occurs in 1 in 20 young men and is idiopathic in origin in most. We have reported that the leucine-rich repeat (LRR) and guanylate kinase-like domain containing, isoform (LRGUK)-1 is essential for sperm head shaping, the manchette, and the initiation of sperm tail growth from the centriole/basal body, and thus, male fertility. Within this study we have used a yeast 2-hybrid screen of an adult testis library to identify LRGUK1-binding partners, which were then validated with a range of techniques. The data indicate that LRGUK1 likely achieves its function in partnership with members of the HOOK family of proteins (HOOK-1-3), Rab3-interacting molecule binding protein (RIMBP)-3 and kinesin light chain (KLC)-3, all of which are associated with intracellular protein transport as cargo adaptor proteins and are localized to the manchette. LRGUK1 consists of 3 domains; an LRR, a guanylate kinase (GUK)-like and an unnamed domain. In the present study, we showed that the GUK-like domain is essential for binding to HOOK2 and RIMBP3, and the LRR domain is essential for binding to KLC3. These findings establish LRGUK1 as a key component of a multiprotein complex with an essential role in microtubule dynamics within haploid male germ cells.-Okuda, H., DeBoer, K., O'Connor, A. E., Merriner, D. J., Jamsai, D., O'Bryan, M. K. LRGUK1 is part of a multiprotein complex required for manchette function and male fertility.
每20名年轻男性中就有1人患有不育症，且大多数病因不明。我们曾报道，富含亮氨酸重复序列（LRR）和鸟苷酸激酶样结构域的异构体（LRGUK）-1对精子头部塑形、精子颈环以及从中心粒/基体起始精子尾部生长至关重要，因此对男性生育能力也至关重要。在本研究中，我们利用酵母双杂交筛选成年睾丸文库来鉴定与LRGUK1结合的蛋白，随后用一系列技术对其进行验证。数据表明，LRGUK1可能与HOOK蛋白家族（HOOK-1-3）、Rab3相互作用分子结合蛋白（RIMBP）-3和驱动蛋白轻链（KLC）-3的成员协同发挥功能，所有这些蛋白作为货物衔接蛋白都与细胞内蛋白质运输相关，并定位于精子颈环。LRGUK1由3个结构域组成：一个LRR结构域、一个类鸟苷酸激酶（GUK）结构域和一个未命名结构域。在本研究中，我们发现类GUK结构域对于与HOOK2和RIMBP3结合至关重要，而LRR结构域对于与KLC3结合至关重要。这些发现表明LRGUK1是一种多蛋白复合物的关键组成部分，在单倍体雄性生殖细胞的微管动力学中发挥重要作用。-奥库达，H.，德布尔，K.，奥康纳，A.E.，梅里纳，D.J.，贾姆赛伊，D.，奥布赖恩，M.K. LRGUK1是精子颈环功能和男性生育所需的多蛋白复合物的一部分。
