Thöny-Meyer L, Kaiser D
Department of Biochemistry, Stanford University School of Medicine, California 94305-5427.
J Bacteriol. 1993 Nov;175(22):7450-62. doi: 10.1128/jb.175.22.7450-7462.1993.
Two Tn5 lac insertions into the Myxococcus genome at sites omega 4414 and omega 4473, which are separated by 550 nucleotides, inactivate fruiting body development. Sporulation is decreased 100- to 10,000-fold. At least two genes, devR and devS, are transcribed in this region, probably as an operon. Expression of devR begins by 6 h after starvation has initiated development. On the basis of their nucleotide sequences, devR and devS are expected to encode proteins of 302 and 214 amino acids, respectively. Dev+ function can be restored by a segment of 7.8 kb cloned from the devRS region of wild-type cells. Two experiments show that devR expression is under strong negative autoregulation. beta-Galactosidase is expressed at a higher level from a transcriptional devR::lacZ fusion when the fused operon is in a dev strain than when it is in the dev/dev+ genetic background of a partial diploid. There is more mRNA accumulation from the devRS region in the dev strain than in a rescued dev/dev+ tandem duplication strain. Sporulation rescue is correlated with some degree of negative autoregulation, even though sporulation is not inversely proportional to beta-galactosidase expression from omega 4414. A second level of regulation is suggested by complementation of dev by dev+ in duplication strains. The expression of devRS, measured by sporulation levels, differs 1,000-fold when devRS+ is moved from a distance of 20 kb to 3 Mb from the mutant devRS locus. Expression of devR is also dependent on the cell density at which development is initiated, a third level of regulation. Multiple levels of regulation suggest that devRS is a switch required to activate completion of aggregation and sporulation.
两个Tn5 lac插入到粘球菌基因组中位于ω4414和ω4473位点处,这两个位点相隔550个核苷酸,它们使子实体发育失活。孢子形成减少了100至10000倍。在该区域至少有两个基因devR和devS被转录,可能作为一个操纵子。饥饿引发发育后6小时开始devR的表达。根据它们的核苷酸序列,预计devR和devS分别编码302和214个氨基酸的蛋白质。野生型细胞devRS区域克隆的一段7.8 kb片段可恢复Dev+功能。两项实验表明devR的表达受到强烈的负自调控。当融合操纵子处于dev菌株中时,转录的devR::lacZ融合体中β-半乳糖苷酶的表达水平高于其处于部分二倍体的dev/dev+遗传背景中时。dev菌株中devRS区域的mRNA积累比挽救后的dev/dev+串联重复菌株中的更多。孢子形成的挽救与某种程度的负自调控相关,尽管孢子形成与来自ω4414的β-半乳糖苷酶表达并非成反比。重复菌株中dev+对dev的互补作用提示了第二种调控水平。当devRS+从突变的devRS位点移动到距离20 kb至3 Mb处时,通过孢子形成水平测量的devRS表达相差1000倍。devR的表达还取决于发育起始时的细胞密度,这是第三种调控水平。多种调控水平表明devRS是激活聚集和孢子形成完成所需的一个开关。