Ueki Toshiyuki, Inouye Sumiko
Department of Biochemistry, Robert Wood Johnson Medical School, 675 Hoes Lane, Piscataway, NJ 08854, USA.
Proc Natl Acad Sci U S A. 2003 Jul 22;100(15):8782-7. doi: 10.1073/pnas.1533026100. Epub 2003 Jul 8.
Myxococcus xanthus exhibits social behavior and multicellular development. FruA is an essential transcription factor for fruiting body development in M. xanthus. In the present study, the upstream promoter region was found to be necessary for the induction of fruA expression during development. A cis-acting element required for the induction was identified and was located between nucleotides -154 and -107 with respect to the transcription initiation site. In addition, it was found that two binding sites exist within this element of the fruA promoter. By using DNA affinity column chromatography containing the cis-acting element, a fruA promoter-binding protein was purified. The purified protein was shown by N-terminal sequence analysis to be identical to MrpC, a protein identified previously by transposon insertion mutagenesis as an essential locus for fruiting body development [Sun, H. & Shi, W. (2001) J. Bacteriol. 183, 4786-4795]. Furthermore, fruA mRNA was not detectable in the mrpC::km strain, demonstrating that MrpC is essential for fruA expression. Moreover, mutational analysis of the binding sites for MrpC in the fruA promoter indicates that binding of MrpC activates transcription of fruA in vivo. This report provides evidence for a direct molecular interaction involved in temporally regulated gene expression in M. xanthus.
黄色黏球菌表现出社会行为和多细胞发育。FruA是黄色黏球菌子实体发育所必需的转录因子。在本研究中,发现上游启动子区域对于发育过程中fruA表达的诱导是必需的。鉴定出了诱导所需的顺式作用元件,其位于相对于转录起始位点的核苷酸-154至-107之间。此外,发现在fruA启动子的该元件内存在两个结合位点。通过使用含有顺式作用元件的DNA亲和柱层析,纯化了一种fruA启动子结合蛋白。通过N端序列分析表明纯化的蛋白与MrpC相同,MrpC是先前通过转座子插入诱变鉴定为子实体发育必需位点的一种蛋白[Sun, H. & Shi, W. (2001) J. Bacteriol. 183, 4786 - 4795]。此外,在mrpC::km菌株中未检测到fruA mRNA,表明MrpC对于fruA表达是必需的。而且,对fruA启动子中MrpC结合位点的突变分析表明,MrpC的结合在体内激活fruA的转录。本报告为黄色黏球菌中参与时间调控基因表达的直接分子相互作用提供了证据。