GroEL is involved in activation of Escherichia coli RNA polymerase devoid of the omega subunit in vivo.
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. E. coli strains lacking omega (omega-less) are viable, but exhibit a slow-growth phenotype. Renaturation of RNA polymerase isolated from an omega-less mutant, in the presence of omega, resulted in maximum recovery of activity. The omega-less RNA polymerase from omega-less strains recruits the chaperonin, GroEL (unlike the wild-type enzyme), suggesting a structural deformity of the mutant enzyme. The GroEL-containing core RNA polymerase interacts efficiently with sigma70 to generate the fully functional holoenzyme. However, when GroEL was removed, the enzyme was irreversibly nonfunctional and was unable to bind to sigma70. The damaged enzyme regained activity after going through a cycle of denaturation and reconstitution in the presence of omega or GroEL. GroES was found to have an inhibitory effect on the core-sigma70 association unlike the omega subunit. The omega subunit may therefore be needed for stabilization of the structure of RNA polymerase.
Mukherjee
K
K
Centre for Cellular and Molecular Biology, Hyderabad, India.
Nagai
H
H
Shimamoto
N
N
Chatterji
D
D
D001426:Bacterial Proteins / Q000378:metabolism*; D018834:Chaperonin 60 / Q000502:physiology*; D012321:DNA-Directed RNA Polymerases / Q000172:deficiency / Q000235:genetics / Q000378:metabolism / Q000502:physiology*; D004789:Enzyme Activation; D004926:Escherichia coli / Q000201:enzymology* / Q000235:genetics / Q000254:growth & development; D011489:Protein Denaturation; D017510:Protein Folding; D012318:RNA Polymerase I / Q000378:metabolism*; D012808:Sigma Factor / Q000378:metabolism
D001426:Bacterial Proteins; D018834:Chaperonin 60; D012808:Sigma Factor; C111232:RNA polymerase omega subunit; C081775:RNA polymerase sigma 70; D012321:DNA-Directed RNA Polymerases; D012318:RNA Polymerase I