Shadel G S, Devine J H, Baldwin T O
Department of Biochemistry and Biophysics, Texas A & M University, College Station 77843.
J Biolumin Chemilumin. 1990 Apr-Jun;5(2):99-106. doi: 10.1002/bio.1170050205.
Regulation of expression of bioluminescence from the Vibrio fischeri lux regulon in Escherichia coli is a consequence of a unique form of positive feedback superimposed on a poorly defined cis-acting repression mechanism. The lux regulon consists of two divergently transcribed operons. The leftward operon contains only a single gene, luxR, which encodes a transcriptional activator protein. The rightward operon contains luxI, which together with luxR and the 218 base pairs separating the two operons comprises the primary regulatory circuit, and the five structural genes, luxC, luxD, luxA, luxB and luxE, which are required for the bioluminescence activity. Transcription of luxR from PL is stimulated by binding of the E. coli crp gene product to the sequence TGTGACAAAAATCCAA upstream of the presumed promoter. Binding of pure E. coli CAP protein in a cAMP-dependent reaction to the V. fischeri lux regulatory region has been demonstrated by in vitro footprinting. The luxI gene product is an enzyme which catalyses a condensation reaction of cytoplasmic substrates to yield the autoinducer, N-(3-oxo-hexanoyl) homoserine lactone. Accumulation of autoinducer, which is freely diffusible, results in formation of a complex with LuxR. The complex binds to the sequence ACCTGTAGGATCGTACAGGT upstream of PR to stimulate transcription of the rightward operon. Increased transcription from PR should yield increased levels of LuxI and higher levels of autoinducer which would further activate LuxR. The LuxR binding site is also a LexA binding site, as demonstrated by in vitro footprinting. Basal transcription from both PL and PR is repressed by sequences within the luxR coding region.(ABSTRACT TRUNCATED AT 250 WORDS)
费氏弧菌lux操纵子在大肠杆菌中生物发光表达的调控,是一种独特的正反馈形式叠加在定义不清的顺式作用阻遏机制上的结果。lux操纵子由两个反向转录的操纵子组成。向左的操纵子仅包含一个基因luxR,它编码一种转录激活蛋白。向右的操纵子包含luxI,luxI与luxR以及分隔两个操纵子的218个碱基对共同构成主要调控回路,还有五个结构基因luxC、luxD、luxA、luxB和luxE,它们是生物发光活性所必需的。来自PL的luxR转录受到大肠杆菌crp基因产物与假定启动子上游序列TGTGACAAAAATCCAA结合的刺激。通过体外足迹法已证明纯大肠杆菌CAP蛋白在依赖cAMP的反应中与费氏弧菌lux调控区域结合。luxI基因产物是一种酶,它催化细胞质底物的缩合反应以产生自诱导物N-(3-氧代己酰基)高丝氨酸内酯。可自由扩散的自诱导物的积累导致与LuxR形成复合物。该复合物与PR上游的序列ACCTGTAGGATCGTACAGGT结合,以刺激向右操纵子的转录。PR转录增加应导致LuxI水平升高和自诱导物水平更高,这将进一步激活LuxR。如体外足迹法所示,LuxR结合位点也是LexA结合位点。来自PL和PR的基础转录受到luxR编码区内序列的抑制。(摘要截短至250字)