Repression of the Escherichia coli modABCD (molybdate transport) operon by ModE.
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 genes codes for a 490-amino-acid protein. ModF (54 kDa). The role of ModF has not yet been determined; however, mutations in modE depressed 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 GITA. A highly conserved amino acid sequence, TSARNOXXG (amino acids 125 to 133), was identified in ModE and homologs from Azotobacter vinelandii, Haemophilus influenzae, Rhodobacter capsulatus, and Clostridium pasterianum. 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.
Grunden
A M
AM
Department of Microbiology and Cell Science, University of Florida, Gainesville 32611, USA.
Ray
R M
RM
Rosentel
J K
JK
Healy
F G
FG
Shanmugam
K T
KT
D000595:Amino Acid Sequence; D001426:Bacterial Proteins / Q000502:physiology*; D001483:Base Sequence; D001665:Binding Sites; D001692:Biological Transport; D004247:DNA / Q000378:metabolism; D004926:Escherichia coli / Q000235:genetics / Q000378:metabolism*; D005809:Genes, Regulator; D008969:Molecular Sequence Data; D008982:Molybdenum / Q000378:metabolism*; D009154:Mutation; D009876:Operon; D012097:Repressor Proteins / Q000502:physiology*
D001426:Bacterial Proteins; D012097:Repressor Proteins; C044659:molybdate; D008982:Molybdenum; D004247:DNA