Kalyuzhnaya Marina G, Lidstrom Mary E
Department of Chemical Engineering. Department of Microbiology, University of Washington, Seattle, Washington 98195-1750, USA.
J Bacteriol. 2003 Feb;185(4):1229-35. doi: 10.1128/JB.185.4.1229-1235.2003.
A new gene, qscR, encoding a LysR-type transcriptional regulator that is a homolog of CbbR, has been characterized from the facultative methylotroph Methylobacterium extorquens AM1 and shown to be the major regulator of the serine cycle, the specific C1 assimilation pathway. The qscR mutant was shown to be unable to grow on C1 compounds, and it lacked the activity of serine-glyoxylate aminotransferase, a key enzyme of the serine cycle. Activities of other serine cycle enzymes were decreased during growth on C1 compounds compared to the activities found in wild-type M. extorquens AM1. Promoter fusion assays, as well as reverse transcription-PCR assays, have indicated that the serine cycle genes belong to three separate transcriptional units, sga-hpr-mtdA-fch, mtkA-mtkB-ppc-mcl, and gly. Gel retardation assays involving the purified QscR have demonstrated the specific binding of QscR to the DNA regions upstream of sga, mtkA, gly, and qscR. We conclude that QscR acts as a positive transcriptional regulator of most of the serine cycle enzymes and also as an autorepressor.
从兼性甲基营养菌嗜甲基甲基杆菌AM1中鉴定出一个新基因qscR,它编码一种LysR型转录调节因子,是CbbR的同源物,并且已证明它是丝氨酸循环(特定的C1同化途径)的主要调节因子。qscR突变体在C1化合物上无法生长,并且缺乏丝氨酸 - 乙醛酸转氨酶(丝氨酸循环的关键酶)的活性。与野生型嗜甲基甲基杆菌AM1相比,在C1化合物上生长期间,其他丝氨酸循环酶的活性降低。启动子融合试验以及逆转录 - PCR试验表明,丝氨酸循环基因属于三个独立的转录单元,即sga - hpr - mtdA - fch、mtkA - mtkB - ppc - mcl和gly。涉及纯化的QscR的凝胶阻滞试验证明了QscR与sga、mtkA、gly和qscR上游的DNA区域特异性结合。我们得出结论,QscR作为大多数丝氨酸循环酶的正转录调节因子,同时也作为一种自动阻遏物。