Interaction surface of bacteriophage P4 protein Psu required for complex formation with the transcription terminator Rho.
Rho-dependent transcription termination is an essential function in prokaryotes, and the transcription terminator Rho is highly conserved among different species. The bacteriophage P4 capsid-decoration protein, Psu, interacts specifically with and inhibits the function of Escherichia coli Rho. The interaction surface of Psu involved in interacting with Rho is not known, but knowledge of this is important to understand the mechanism of its action and will be useful to design peptide inhibitor(s) for Rho. We have isolated and characterized seven Psu mutants defective in interacting with Rho and in exerting anti-Rho activity. Conformational probing of Psu revealed that the N-terminal region of the protein folds over onto its central part, forming a globular domain and leaving a solvent-exposed "tail" in the C-terminus. The mutations are located in both of these domains. N-terminal mutants are instrumental in disrupting the N- to C-terminal "cross-talk" in Psu that is required for its structural integrity and its function. Site-specific cross-linking experiments showed that the C-terminal tail preferentially cross-links to Rho and this region of Psu is protected from limited proteolysis when bound to Rho. Therefore, the mutations in this region may have affected the direct interaction of Psu with Rho. We propose that the globular N-terminal domain of Psu confers structural integrity to the functionally important C-terminal tail, which interacts directly with the hexameric Rho.
Pani
Bibhusita
B
Laboratory of Transcription Biology, Centre for DNA Fingerprinting and Diagnostics, Tuljaguda, Nampally, Hyderabad-500001, India.
Ranjan
Amitabh
A
Sen
Ranjan
R
D000595:Amino Acid Sequence; D036022:Capsid Proteins / Q000737:chemistry / Q000235:genetics / Q000378:metabolism*; D003432:Cross-Linking Reagents / Q000737:chemistry; D008969:Molecular Sequence Data; D009154:Mutation; D011487:Protein Conformation; D012234:Rho Factor / Q000235:genetics / Q000378:metabolism*; D013728:Terminator Regions, Genetic; D014158:Transcription, Genetic
D036022:Capsid Proteins; D003432:Cross-Linking Reagents; D012234:Rho Factor; C074513:polarity suppression factor, bacteriophage P4