Repression of the Escherichia coli modABCD (molybdate transport) operon by ModE

被引:91
作者
Grunden, AM [1 ]
Ray, RM [1 ]
Rosentel, JK [1 ]
Healy, FG [1 ]
Shanmugam, KT [1 ]
机构
[1] UNIV FLORIDA, DEPT MICROBIOL & CELL SCI, GAINESVILLE, FL 32611 USA
关键词
D O I
10.1128/jb.178.3.735-744.1996
中图分类号
Q93 [微生物学];
学科分类号
071005 ; 100705 ;
摘要
The modABC gene products constitute the molybdate-specific transport system in Escherichia coli. Another operon coding for two proteins which diverges from the modABCD operon has been identified. The first gene of this operon codes for a 262-amino-acid protein, designated ModE (28 kDa), and the second gene codes for a 490-amino-acid protein, ModF (54 kDa). The role of ModF has not yet been determined; however, mutations in modE derepressed modABCD transcription even in the presence of molybdate, suggesting that ModE is a repressor. ModE, in the presence of 1 mM molybdate, repressed the production of plasmid-encoded ModA and ModB' proteins in an in vitro transcription-translation system. DNA mobility shift experiments confirmed that ModE binds to an oligonucleotide derived from the operator region of the modABCD operon. Further experimentation indicated that ModE binding to target DNA minimally requires an 8-bp inverted-repeat sequence, TAAC.GTTA. A highly conserved amino acid sequence, TSARNQXXG (amino acids 125 to 133), was identified in ModE and homologs from Azotobacter vinelandii, Haemophilus influenzae, Rhodobacter capsulatus, and Clostridium pasteurianum. Mutants with mutations in either T or G of this amino acid sequence were isolated as ''superrepressor'' mutants. These mutant proteins repressed modABCD transcription even in the absence of molybdate, which implies that this stretch of amino acids is essential for the binding of molybdate by the ModE protein. These results show that molybdate transport in E. coli is regulated by ModE, which acts as a repressor when bound to molybdate.
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页码:735 / 744
页数:10
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