Studies on the omega subunit of Escherichia coli RNA polymerase--its role in the recovery of denatured enzyme activity.
Highly purified Escherichia coli RNA polymerase contains a small subunit termed omega that has a molecular mass of 10,105 Da and is comprised of 91 amino acids. To elucidate the function of omega, whose role is as yet undefined, the subunit was purified to over 95% purity from an overproducing strain [BL21 (pGP1-2, pE3C-2)]. Purified omega was then reconstituted with RNA polymerase isolated from an omega-less mutant. Externally added omega inhibited promoter-specific transcriptional activity at all promoters tested. Renaturation of fully denatured omega-less RNA polymerase in the presence of excess omega yielded maximum recovery of activity suggesting a structural rather than functional role for omega.
Mukherjee
K
K
Centre for Cellular and Molecular Biology, Hyderabad, India.
Chatterji
D
D
D012321:DNA-Directed RNA Polymerases / Q000737:chemistry / Q000302:isolation & purification / Q000378:metabolism*; D004926:Escherichia coli / Q000201:enzymology*; D011401:Promoter Regions, Genetic; D011489:Protein Denaturation; D014158:Transcription, Genetic
D012321:DNA-Directed RNA Polymerases