同源染色体联会时形成的联会复合体(Synaptonemal complex, SC)是由减数分裂前期Ⅰ多种蛋白质聚集而成的超级复合结构。生殖细胞特异性的核蛋白C(2)M(Crossover suppressor on 2 of Manheim)在染色体上高度聚集可以诱导SC的形成。本文采用酵母双杂交方法,利用C(2)M的诱饵表达载体筛选果蝇cDNA文库,共发现40个可能与C(2)M相互作用的蛋白,包括多种DNA及组蛋白结合蛋白、ATPase、转录调节因子。从筛选的结果中,选取wech和Psf1基因构建了转基因果蝇,并在生殖细胞中进行了基因沉默,结果显示联会复合体的消失受到延迟。上述结果表明Wech和Psf1蛋白可能与C(2)M形成复合物,共同参与联会复合体的形成或其稳定性的维持。
The synaptonemal complex (SC) is a huge structure which assembles between the homologous chromosomes during meiotic prophase I. Drosophila germ cell-specific nucleoprotein C(2)M clustering at chromosomes can induce SC formation. To further study the molecular function and mechanism of C(2)M in meiosis, we constructed a bait vector for C(2)M and used the yeast two-hybrid system to identify C(2)M interacting proteins. Forty interacting proteins were obtained, including many DNA and histone binding proteins, ATP synthases and transcription factors. Gene silencing assays in Drosophila showed that two genes, wech and Psf1, may delay the disappearance of SC. These results indicate that Wech and Psf1 may form a complex with C(2)M to participate in the formation or stabilization of the SC complex.
[1] Tanneti NS, Landy K, Joyce EF, McKim KS. A pathway for synapsis initiation during zygotene in Drosophila Oocytes. Curr Biol , 2011, 21(21): 1852-1857.
[2] Balicky EM, Endres MW, Lai C, Bickel SE. Meiotic cohesion requires accumulation of ORD on chromosomes before condensation. Mol Biol Cell , 2002, 13(11): 3890-3900.
[3] Anderson LK, Royer SM, Page SL, McKim KS, Lai A, Lily MA, Hawley RS. Juxtaposition of C(2)M and the transverse filament protein C(3)G within the central region of Drosophila synaptonemal complex. Proc Natl Acad Sci USA , 2005, 102(12): 4482-4487.
[4] Lake CM, Hawley RS. The molecular control of meiotic chromosomal behavior: events in early meiotic prophase in Drosophila Oocytes. Annu Rev Physiol , 2012, 74: 425-451.
[5] Heidmann D, Horn S, Heidmann S, Schleiffer A, Nasmyth K, Lehner CF. The Drosophila meiotic kleisin C(2)M functions before the meiotic divisions. Chromosoma , 2004, 113(4): 177-187.
[6] Manheim EA, McKim KS. The synaptonemal complex component C(2)M regulates meiotic crossing over in Drosophila . Curr Biol , 2003, 13(4): 276-285.
[7] Fields S, Song O. A novel genetic system to detect protein- protein interactions. Nature , 1989, 340(6230): 245-246.
[8] 杨姗姗, 孙晓丽, 于洋, 才华, 纪巍, 柏锡, 朱延明. 酵母双杂交筛选与GsCBRLK相互作用的蛋白质. 遗传, 2013, 35(3): 388-394.
[9] 都建, 陈立建, 沈继龙, 花沙沙, 姚雪彪. 用酵母双杂交系统筛选CENP2E相互作用蛋白. 中国生物化学与分子生物学报, 2009, 25(8): 719-726.
[10] Lee JY, Orr-Weaver TL. The molecular basis of sister chromatid cohesion. Annu Rev Cell Dev Biol , 2001, 17: 753-777.
[11] Ono T, Losada A, Hirano M, Myers MP, Neuwald AF, Hirano T. Differential contributions of condensinⅠand condensinⅡ to mitotic chromosome architecture in vertebrate cells. Cell , 2003, 115(1): 109-121.
[12] Dej KJ, Ahn C, Orr-Weaver TL. Mutations in the Drosophila condensin subunit dCAP-G: defining the role of condensin for chromosome condensation in mitosis and gene expression in interphase. Genetics , 2004, 168(2): 895-906.
[13] Herzog S, Jaiswal SN, Urban E, Riemer A, Fischer S, Heidmann SK. Functional dissection of the Drosophila melanogaster condensin subunit Cap-G reveals its exclusive association with condensinⅠ. PLoS Genet , 2013, 9(4): e1003463.
[14] Meroni G, Diez-Roux G. TRIM/RBCC, a novel class of “single protein Ring finger” E3 ubiquitin ligases. BioEssays , 2005, 27(11): 1147-1157.
[15] Löer B, Bauer R, Bornheim R, Grell J, Kremmer E, Kolanus W, Hoch M. The NHL-domain protein Wech is crucial for the integrin-cytoskeleton link. Nat Cell Biol , 2008, 10(4): 422-428.
[16] Löer B, Hoch M. Wech proteins: roles in integrin functions and beyond. Cell Adhes Migr , 2008, 2(3): 177-179.
[17] Li Y, Zhang Q, Carreira-Rosario A, Maines JZ, McKearin DM, Buszczak M. Mei-p26 cooperates with Bam, Bgcn and Sxl to promote early germline development in the Drosophila ovary. PLoS One , 2013, 8(3): e58301.
[18] Lee CY, Wilkinson BD, Siegrist SE, Wharton RP, Doe CQ. Brat is a Miranda cargo protein that promotes neuronal differentiation and inhibits neuroblast self-renewal. Dev Cell , 2006, 10(4): 441-449.
[19] Arama E, Dickman D, Kimchie Z, Shearn A, Lev Z. Mutations in the β-propeller domain of the Drosophila brain tumor ( brat ) protein induce neoplasm in the larval brain. Oncogene , 2000, 19(33): 3706-3716.
[20] Grabowska E, Wronska U, Denkiewicz M, Jaszczur M, Respondek A, Alabrudzinska M, Suski C, Makiela-Dzbenska K, Jonczyk P, Fijalkowska IJ. Proper functioning of the GINS complex is important for the fidelity of DNA replication in yeast. Mol Microbiol , 2014, 92(4): 659-680.
