The Escherichia coli rpoB60 mutation blocks antitermination by coliphage HK022 Q-function.
The lambdoid bacteriophage regulate gene expression by suppressing transcription terminators. Although similar in sequence to lambda, HK022 lacks an analogue to the lambda N antitermination gene and a distinct nutR sequence. To define the HK022 antitermination system, we plated the phage on Escherichia coli nus mutants that inhibit lambda N function. Only rpoB60 (also called nusC60) blocked HK022 lytic growth. Analyses of HK022-lambda hybrid phage suggested that a HK022 function analogous to lambda Q was inhibited by rpoB60. This result was confirmed with pR'-tR'-galK fusions. HK022 Q-protein suppressed tR' in wild-type but not in rpoB60 mutants. The lambda Q-protein, although inhibited by rpoB60, was more active than the HK022 analogue. A single amino acid difference between the two Q-proteins accounts for the phenotype. Changing the penultimate residue of HK022 Q from alanine to the lambda threonine generated a phage that could propagate on rpoB60 hosts. Host and phage mutations that permitted HK022 growth in rpoB60 strains were characterized. The bacterial suppressors were located in the Escherichia coli nusB gene. The phage suppressors represented recessive mutations in a HK022 b-region sequence encoding an open reading frame of 73 codons.
Atkinson
B L
BL
Institute of Cancer Research, Columbia University, College of Physicians and Surgeons, New York, NY 10032.
Gottesman
M E
ME
D000595:Amino Acid Sequence; D001483:Base Sequence; D003090:Coliphages / Q000235:genetics*; D004926:Escherichia coli / Q000235:genetics*; D015967:Gene Expression Regulation, Viral; D016153:Genes, Suppressor; D005814:Genes, Viral; D005816:Genetic Complementation Test; D008969:Molecular Sequence Data; D009154:Mutation; D013728:Terminator Regions, Genetic; D014158:Transcription, Genetic; D015678:Viral Structural Proteins / Q000235:genetics*
D015678:Viral Structural Proteins