In vivo and in vitro evidence for an anti-Rho activity induced by the phage P4 polarity suppressor protein Psu.
The polarity suppression (Psu) protein of bacteriophage P4 causes suppression of transcriptional polarity in Escherichia coli by overcoming Rho termination factor activity. Two new psu mutants defective in polarity suppression are described. The psu5 mutation deletes codons 95-98 from about the middle of the gene, and the mutant protein is inactive. The psu6 mutation changes Phe169 to Val and encodes a temperature-sensitive protein. Constitutive overexpression of psu+ from a plasmid prevents colony formation, but overexpression of mutant genes (psu5, psu6) does not, suggesting that Psu disturbs essential host function(s). Rho protein synthesis is enhanced several-fold in cells containing wild-type Psu, due to readthrough at the rho attenuator, while the physical stability of Rho is maintained. As a consequence, Psu-producing cells accumulate significantly more Rho than normal cells, reminiscent of termination-defective rho mutants. The polarity suppression activity induced by Psu is demonstrated in vitro by the efficient readthrough of Rho-dependent terminators lambda tR1 and TIS2 during coupled transcription-translation. Purified Rho protein restores termination at TIS2 when added to Psu-containing reactions but NusG does not. The data support the hypothesis that Psu has or elicits an anti-Rho function.
Linderoth
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Department of Molecular and Cell Biology, University of California, Berkeley, California, 94720-3202, USA.
Tang
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Calendar
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D002213:Capsid / Q000235:genetics / Q000502:physiology*; D036022:Capsid Proteins; D003090:Coliphages / Q000502:physiology*; D004926:Escherichia coli / Q000235:genetics / Q000821:virology; D009154:Mutation; D014176:Protein Biosynthesis; D012234:Rho Factor / Q000037:antagonists & inhibitors* / Q000096:biosynthesis / Q000302:isolation & purification / Q000378:metabolism; D013728:Terminator Regions, Genetic; D014158:Transcription, Genetic
D036022:Capsid Proteins; D012234:Rho Factor; C074513:polarity suppression factor, bacteriophage P4