Margalit Ayelet, Segura-Totten Miriam, Gruenbaum Yosef, Wilson Katherine L
Department of Genetics, Institute of Life Sciences, Hebrew University of Jerusalem, Givat Ram, Jerusalem 91904, Israel.
Proc Natl Acad Sci U S A. 2005 Mar 1;102(9):3290-5. doi: 10.1073/pnas.0408364102. Epub 2005 Feb 22.
Barrier-to-autointegration factor (BAF) binds dsDNA, LEM-domain proteins, and lamins. Caenorhabditis elegans BAF requires Ce-lamin and two LEM-domain proteins (Ce-emerin and Ce-MAN1) to localize during nuclear assembly. It was unknown whether Ce-lamin and LEM proteins, in turn, depend on Ce-BAF (mutually dependent structural roles). RNA interference-mediated down-regulation of Ce-BAF caused gross defects in chromosome segregation, chromatin decondensation, and mitotic progression as early as the two-cell stage, and embryos died at the approximately 100-cell stage. Nuclear pores reassembled, whereas Ce-lamin, Ce-emerin, and Ce-MAN1 bound chromatin but remained patchy and disorganized. The nuclear membranes formed but failed to enclose anaphase-bridged chromatin. Time-lapse imaging showed two phenotypes: anaphase-bridged chromatin that eventually resolved, and segregated chromatin that returned to the midzone. Thus, the assembly of BAF, lamins, and LEM-domain proteins is mutually dependent, and is required to capture segregated chromosomes within the nascent nuclear envelope. Embryos that escaped lethality by down-regulation of Ce-BAF grew into sterile adults with misplaced distal tip cells and gonads, further suggesting that mild postembryonic reductions in BAF disrupt tissue-specific functions.
自身整合屏障因子(BAF)可结合双链DNA、LEM结构域蛋白和核纤层蛋白。秀丽隐杆线虫的BAF在核组装过程中需要Ce-核纤层蛋白以及两种LEM结构域蛋白(Ce-emerin和Ce-MAN1)来定位。此前尚不清楚Ce-核纤层蛋白和LEM蛋白反过来是否依赖于Ce-BAF(相互依赖的结构作用)。RNA干扰介导的Ce-BAF下调早在二细胞阶段就导致染色体分离、染色质解聚和有丝分裂进程出现严重缺陷,胚胎在大约100细胞阶段死亡。核孔重新组装,而Ce-核纤层蛋白、Ce-emerin和Ce-MAN1与染色质结合,但仍呈斑片状且无序。核膜形成,但未能包围后期桥接的染色质。延时成像显示出两种表型:最终得以解决的后期桥接染色质,以及回到中区的分离染色质。因此,BAF、核纤层蛋白和LEM结构域蛋白的组装是相互依赖的,并且是在新生核膜内捕获分离染色体所必需的。通过下调Ce-BAF而逃过致死性的胚胎发育成不育成虫,其远端末梢细胞和性腺位置异常,这进一步表明BAF在胚胎后期的轻度减少会破坏组织特异性功能。