Krause M, Harrison S W, Xu S Q, Chen L, Fire A
Laboratory of Molecular Biology, National Institute of Diabetes, Digestive, and Kidney Diseases, NIH, Bethesda, Maryland 20892.
Dev Biol. 1994 Nov;166(1):133-48. doi: 10.1006/dbio.1994.1302.
We investigated the cis-acting sequences regulating expression of the Caenorhabditis elegans gene hlh-1, a homolog of the MyoD family of myogenic regulatory factors. The hlh-1 gene is expressed in mature body wall muscle, in clonal muscle precursors, in a set of early embryonic blastomeres (the MS-granddaughters), and in six glial-like cells called GLRs. The entire natural hlh-1 expression pattern is recapitulated in transgenic animals containing an hlh-1::lacZ fusion with 5.1 kb of hlh-1 sequences beginning upstream of the coding region and extending into the second exon. Deletions and rearrangements in this 5.1-kb sequence were assayed for their effects on reporter gene expression in transgenic animals. Deletions removing a segment 434-550 bp upstream of the coding region resulted in a loss of all aspects of the expression pattern, suggesting that at least one common regulatory factor is required for all expression. Deletions of other regulatory elements affected distinct aspects of the expression pattern; hence, each expression subpattern exhibits a different set of sequence requirements. Interspecies sequence comparison and lacZ expression constructs with the related nematode Caenorhabditis briggsae indicated that the C. briggsae and C. elegans genes contain equivalent sets of control signals. The results from this work implicate the hlh-1 promoter as an integration point for diverse temporal and spatial control signals.
我们研究了调控秀丽隐杆线虫基因hlh-1表达的顺式作用序列,hlh-1是肌源性调节因子MyoD家族的一个同源基因。hlh-1基因在成熟的体壁肌肉、克隆性肌肉前体细胞、一组早期胚胎卵裂球(MS的孙女细胞)以及六个称为GLR的神经胶质样细胞中表达。在含有hlh-1::lacZ融合基因的转基因动物中重现了整个天然hlh-1的表达模式,该融合基因带有从编码区上游开始并延伸至第二个外显子的5.1 kb的hlh-1序列。检测了该5.1 kb序列中的缺失和重排对转基因动物中报告基因表达的影响。缺失编码区上游434 - 550 bp的片段导致表达模式的所有方面丧失,这表明所有表达至少需要一种共同的调节因子。其他调节元件的缺失影响了表达模式的不同方面;因此,每个表达亚模式表现出不同的一组序列要求。与相关线虫秀丽新杆线虫进行种间序列比较和lacZ表达构建体表明,秀丽新杆线虫和秀丽隐杆线虫的基因包含等效的控制信号集。这项工作的结果表明hlh-1启动子是多种时间和空间控制信号的整合点。