Sequence and characterization of Bacillus subtilis CheW.
A Bacillus subtilis open reading frame (ORF) encoding a predicted polypeptide of 156 amino acids was subcloned and sequenced. The polypeptide was found to be homologous to CheW of Escherichia coli, sharing 28.6% amino acid identity. The ORF was verified by using a bacteriophage T7 expression system in E. coli. The gene was inactivated by insertion of a nonpolor chloramphenicol acetyltransferase cassette in its N-terminal region. In the absence of chemoeffectors, the mutant displayed a smooth swimming bias, with some tumbling. The CheW- mutant was defective on swarm plates but was complemented by a plasmid that expressed wild type CheW. Addition of attractant or repellent to the CheW- mutant resulted in transient smooth swimming or tumbling, respectively. However, capillary assays revealed that chemotaxis was substantially impaired in the mutant strain.
Hanlon
D W
DW
Department of Biochemistry, College of Medicine, University of Illinois, Urbana 61801.
Márquez-Magaña
L M
LM
Carpenter
P B
PB
Chamberlin
M J
MJ
Ordal
G W
GW
D000595:Amino Acid Sequence; D001224:Aspartic Acid / Q000378:metabolism; D001412:Bacillus subtilis / Q000235:genetics*; D001426:Bacterial Proteins / Q000235:genetics*; D001483:Base Sequence; D015139:Blotting, Southern; D002630:Chemotactic Factors / Q000235:genetics*; D015500:Chloramphenicol O-Acetyltransferase / Q000235:genetics; D004269:DNA, Bacterial / Q000235:genetics; D003847:Deoxyglucose / Q000378:metabolism; D004591:Electrophoresis, Polyacrylamide Gel; D029968:Escherichia coli Proteins; D005798:Genes, Bacterial; D005816:Genetic Complementation Test; D008969:Molecular Sequence Data; D016366:Open Reading Frames; D010957:Plasmids; D015183:Restriction Mapping; D016415:Sequence Alignment
D001426:Bacterial Proteins; C489103:CheW protein, E coli; D002630:Chemotactic Factors; D004269:DNA, Bacterial; D029968:Escherichia coli Proteins; C051236:CheW protein, Bacteria; D001224:Aspartic Acid; D003847:Deoxyglucose; D015500:Chloramphenicol O-Acetyltransferase