Kraemer B, Crittenden S, Gallegos M, Moulder G, Barstead R, Kimble J, Wickens M
Department of Biochemistry University of Wisconsin-Madison Madison, Wisconsin 53706, USA.
Curr Biol. 1999 Sep 23;9(18):1009-18. doi: 10.1016/s0960-9822(99)80449-7.
The Caenorhabditis elegans FBF protein and its Drosophila relative, Pumilio, define a large family of eukaryotic RNA-binding proteins. By binding regulatory elements in the 3' untranslated regions (UTRs) of their cognate RNAs, FBF and Pumilio have key post-transcriptional roles in early developmental decisions. In C. elegans, FBF is required for repression of fem-3 mRNA to achieve the hermaphrodite switch from spermatogenesis to oogenesis.
We report here that FBF and NANOS-3 (NOS-3), one of three C. elegans Nanos homologs, interact with each other in both yeast two-hybrid and in vitro assays. We have delineated the portions of each protein required for this interaction. Worms lacking nanos function were derived either by RNA-mediated interference (nos-1 and nos-2) or by use of a deletion mutant (nos-3). The roles of the three nos genes overlap during germ-line development. In certain nos-deficient animals, the hermaphrodite sperm-oocyte switch was defective, leading to the production of excess sperm and no oocytes. In other nos-deficient animals, the entire germ line died during larval development. This germ-line death did not require CED-3, a protease required for apoptosis.
The data suggest that NOS-3 participates in the sperm-oocyte switch through its physical interaction with FBF, forming a regulatory complex that controls fem-3 mRNA. NOS-1 and NOS-2 also function in the switch, but do not interact directly with FBF. The three C. elegans nanos genes, like Drosophila nanos, are also critical for germ-line survival. We propose that this may have been the primitive function of nanos genes.
秀丽隐杆线虫的FBF蛋白及其果蝇同源物Pumilio,定义了一个大型的真核RNA结合蛋白家族。通过结合其同源RNA的3'非翻译区(UTR)中的调控元件,FBF和Pumilio在早期发育决策中具有关键的转录后作用。在秀丽隐杆线虫中,抑制fem-3 mRNA以实现雌雄同体从精子发生到卵子发生的转变需要FBF。
我们在此报告,FBF和秀丽隐杆线虫三个Nanos同源物之一的NANOS-3(NOS-3),在酵母双杂交和体外实验中相互作用。我们已经确定了这种相互作用所需的每种蛋白质的部分。缺乏nanos功能的线虫要么通过RNA介导的干扰(nos-1和nos-2)获得,要么通过使用缺失突变体(nos-3)获得。这三个nos基因在生殖系发育过程中的作用重叠。在某些nos缺陷动物中,雌雄同体的精子-卵子转变存在缺陷,导致产生过多精子且没有卵子。在其他nos缺陷动物中,整个生殖系在幼虫发育期间死亡。这种生殖系死亡不需要CED-3,CED-3是细胞凋亡所需的一种蛋白酶。
数据表明,NOS-3通过与FBF的物理相互作用参与精子-卵子转变,形成一个控制fem-3 mRNA的调控复合物。NOS-1和NOS-2在该转变中也起作用,但不直接与FBF相互作用。秀丽隐杆线虫的三个nanos基因,与果蝇的nanos一样,对生殖系存活也至关重要。我们提出,这可能是nanos基因的原始功能。