Appleford Peter J, Gravato-Nobre Maria, Braun Toby, Woollard Alison
Genetics Unit, Department of Biochemistry, University of Oxford, South Parks Road, Oxford OX1 3QU, UK.
Dev Biol. 2008 May 15;317(2):695-704. doi: 10.1016/j.ydbio.2008.02.049. Epub 2008 Mar 18.
We have identified Conserved Non-coding Elements (CNEs) in the regulatory region of Caenorhabditis elegans and Caenorhabditis briggsae mab-9, a T-box gene known to be important for cell fate specification in the developing C. elegans hindgut. Two adjacent CNEs (a region 78 bp in length) are both necessary and sufficient to drive reporter gene expression in posterior hypodermal cells. The failure of a genomic mab-9::gfp construct lacking this region to express in posterior hypodermis correlates with the inability of this construct to completely rescue the mab-9 mutant phenotype. Transgenic males carrying this construct in a mab-9 mutant background exhibit tail abnormalities including morphogenetic defects, altered tail autofluorescence and abnormal lectin-binding properties. Hermaphrodites display reduced susceptibility to the C. elegans pathogen Microbacterium nematophilum. This comparative genomics approach has therefore revealed a previously unknown role for mab-9 in hypodermal function and we suggest that MAB-9 is required for the secretion and/or modification of posterior cuticle.
我们已在秀丽隐杆线虫和布里格氏秀丽隐杆线虫mab-9(一种T-box基因,已知对秀丽隐杆线虫发育中的后肠细胞命运特化很重要)的调控区域中鉴定出保守非编码元件(CNEs)。两个相邻的CNEs(一个长度为78 bp的区域)对于驱动报告基因在后皮下细胞中的表达既必要又充分。缺乏该区域的基因组mab-9::gfp构建体在后皮下无法表达,这与该构建体无法完全挽救mab-9突变体表型相关。在mab-9突变背景下携带该构建体的转基因雄性表现出尾部异常,包括形态发生缺陷、尾部自发荧光改变和异常的凝集素结合特性。雌雄同体对秀丽隐杆线虫病原体嗜线虫微杆菌的易感性降低。因此,这种比较基因组学方法揭示了mab-9在皮下功能中以前未知的作用,我们认为MAB-9是后表皮分泌和/或修饰所必需的。