Extragenic suppression of motA missense mutations of Escherichia coli.
The MotA and MotB proteins are thought to comprise elements of the stator component of the flagellar motor of Escherichia coli. In an effort to understand interactions among proteins within the motor, we attempted to identify extragenic suppressors of 31 dominant, plasmid-borne alleles of motA. Strains containing these mutations were either nonmotile or had severely impaired motility. Four of the mutants yielded extragenic suppressors mapping to the FlaII or FlaIIIB regions of the chromosome. Two types of suppression were observed. Suppression of one type (class I) probably results from increased expression of the chromosomal motB gene due to relief of polarity. Class I suppressors were partial deletions of Mu insertion sequences in the disrupted chromosomal motA gene. Class I suppression was mimicked by expressing the wild-type MotB protein from a second, compatible plasmid. Suppression of the other type (class II) was weaker, and it was not mimicked by overproduction of wild-type MotB protein. Class II suppressors were point mutations in the chromosomal motB or fliG genes. Among 14 independent class II suppressors characterized by DNA sequencing, we identified six different amino acid substitutions in MotB and one substitution in FliG. A number of the strongest class II suppressors had alterations of residues 136 to 138 of MotB. This particular region within the large, C-terminal periplasmic domain of MotB has previously not been associated with a specific function. We suggest that residues 136 to 138 of MotB may interact directly with the periplasmic face of MotA or help position the N-terminal membrane-spanning helix of MotB properly to interact with the membrane-spanning helices of the MotA proton channel.
Department of Biology, Texas A&M University, College Station 77843-3258, USA.
A G
AG
Garza
P A
PA
Bronstein
P A
PA
Valdez
L W
LW
Harris-Haller
M D
MD
Manson
D000483:Alleles; D001426:Bacterial Proteins / Q000235:genetics*; D002874:Chromosome Mapping; D004706:Endodeoxyribonucleases; D004926:Escherichia coli / Q000235:genetics*; D029968:Escherichia coli Proteins; D015964:Gene Expression Regulation, Bacterial; D013489:Suppression, Genetic
D001426:Bacterial Proteins; D029968:Escherichia coli Proteins; C075660:Flig protein, Bacteria; C063010:MotA protein, Bacteria; C064779:MotB protein, Bacteria; D004706:Endodeoxyribonucleases; C066567:UvrC protein, E coli