PMID:12421556
| Citation |
Nickels, BE, Dove, SL, Murakami, KS, Darst, SA and Hochschild, A (2002) Protein-protein and protein-DNA interactions of sigma70 region 4 involved in transcription activation by lambdacI. J. Mol. Biol. 324:17-34 |
|---|---|
| Abstract |
The cI protein of bacteriophage lambda (lambdacI) activates transcription from promoter P(RM) through an acidic patch on the surface of its DNA-binding domain. Genetic evidence suggests that this acidic patch stimulates transcription from P(RM) through contact with the C-terminal domain (region 4) of the sigma(70) subunit of Escherichia coli RNA polymerase. Here, we identify two basic residues in region 4 of sigma(70) that are critical for lambdacI-mediated activation of transcription from P(RM). On the basis of structural modeling, we propose that one of these sigma(70) residues, K593, facilitates the interaction between lambdacI and region 4 of sigma(70) by inducing a bend in the DNA upstream of the -35 element, whereas the other, R588, interacts directly with a critical acidic residue within the activating patch of lambdacI. Residue R588 of sigma(70) has been shown to play an important role in promoter recognition; our findings suggest that the R588 side-chain has a dual function at P(RM), facilitating the interaction of region 4 with the promoter -35 element and participating directly in the protein-protein interaction with lambdacI. |
| Links | |
| Keywords |
Amino Acid Sequence; Amino Acid Substitution; Bacterial Proteins/genetics; Bacterial Proteins/metabolism; Bacteriophage lambda/genetics; Binding Sites; DNA-Binding Proteins/chemistry; DNA-Binding Proteins/genetics; DNA-Binding Proteins/metabolism; DNA-Directed RNA Polymerases/genetics; DNA-Directed RNA Polymerases/metabolism; Kinetics; Models, Molecular; Molecular Biology/methods; Molecular Sequence Data; Promoter Regions, Genetic; Protein Conformation; Repressor Proteins/chemistry; Repressor Proteins/genetics; Repressor Proteins/metabolism; Sigma Factor/genetics; Sigma Factor/metabolism; Thermus/genetics; Transcriptional Activation; Viral Proteins; Viral Regulatory and Accessory Proteins |
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