Thatcher J D, Fernandez A P, Beaster-Jones L, Haun C, Okkema P G
Department of Biological Sciences (M/C 567), University of Illinois at Chicago, 900 S. Ashland Avenue, Chicago, Illinois 60607, USA.
Dev Biol. 2001 Jan 15;229(2):480-93. doi: 10.1006/dbio.2000.9978.
Gene expression in the Caenorhabditis elegans pharynx is regulated in part by organ-specific signals, which in the myo-2 gene target a regulatory sequence called the C sub-element. C sub-element activity requires the organ specification factor PHA-4, a winged-helix transcription factor expressed in all pharyngeal cells. To identify additional factors involved in pharyngeal organogenesis, we performed a yeast one-hybrid screen for C sub-element binding proteins. Here we describe the novel factor PEB-1, which is coexpressed with PHA-4 in many pharyngeal cell types, including muscles, epithelial cells, marginal cells, and glands, but is undetectable in the pharyngeal nervous system. PEB-1 is also detected outside the pharynx in cells surrounding the rectum and vulva, as well as in the germ line. Reduction of peb-1 function using RNAi results in morphological defects in the somatic tissues in which peb-1 is expressed. We have mapped the PEB-1 DNA-binding domain to a 158-residue region, which is unrelated to known DNA-binding proteins but shares some sequence similarity to the Drosophila Mod(mdg4) proteins. PEB-1 specifically recognizes a site in the C subelement that partially overlaps the PHA-4 binding site. Both the PEB-1 and the PHA-4 binding sites are necessary for strong C sub-element enhancer activity in some cells in which these factors are coexpressed. In contrast the PEB-1 site is dispensable for C sub-element activity in pharyngeal neurons. We propose that PEB-1 functions with PHA-4 to activate target gene expression in cells in which they are coexpressed.
秀丽隐杆线虫咽部的基因表达部分受器官特异性信号调控,这些信号在肌动蛋白-2基因中靶向一个称为C亚元件的调控序列。C亚元件的活性需要器官特异性因子PHA-4,这是一种在所有咽部细胞中表达的翼状螺旋转录因子。为了鉴定参与咽部器官发生的其他因子,我们对C亚元件结合蛋白进行了酵母单杂交筛选。在此,我们描述了新因子PEB-1,它在许多咽部细胞类型中与PHA-4共表达,包括肌肉、上皮细胞、边缘细胞和腺体,但在咽部神经系统中检测不到。在直肠和外阴周围的细胞以及生殖系中也能检测到PEB-1在咽部之外的表达。使用RNA干扰降低peb-1的功能会导致peb-1表达的体细胞组织出现形态缺陷。我们已将PEB-1的DNA结合结构域定位到一个158个残基的区域,该区域与已知的DNA结合蛋白无关,但与果蝇Mod(mdg4)蛋白有一些序列相似性。PEB-1特异性识别C亚元件中与PHA-4结合位点部分重叠的一个位点。在这些因子共表达的一些细胞中,PEB-1和PHA-4结合位点对于强大的C亚元件增强子活性都是必需的。相比之下,PEB-1位点对于咽部神经元中C亚元件的活性是可有可无的。我们提出,PEB-1与PHA-4共同作用,在它们共表达的细胞中激活靶基因表达。