Bipartite functional map of the E. coli RNA polymerase alpha subunit: involvement of the C-terminal region in transcription activation by cAMP-CRP.
The alpha subunit of Escherichia coli RNA polymerase plays a major role in the subunit assembly. Carboxyterminal deletion derivatives lacking 73 or 94 amino acid residues were assembled in vitro into enzyme molecules. Core enzymes consisting of these C-terminal-truncated alpha subunits were as active in RNA synthesis as native core enzyme. By the addition of sigma 70 subunit, these mutant enzymes initiated transcription from certain promoters. The mutant RNA polymerases, however, did not show cAMP-CRP activated transcription. These results demonstrate that the N-terminal region of the alpha subunit is involved in the formation of active enzyme molecule, while the C-terminal region plays an essential role in response to transcription activation by cAMP-CRP.
Igarashi
K
K
Department of Molecular Genetics, National Institute of Genetics, Mishima, Shizuoka, Japan.
Ishihama
A
A
D003001:Cloning, Molecular; D000242:Cyclic AMP / Q000502:physiology*; D004252:DNA Mutational Analysis; D012321:DNA-Directed RNA Polymerases / Q000378:metabolism*; D004926:Escherichia coli / Q000201:enzymology; D015964:Gene Expression Regulation, Bacterial; D046911:Macromolecular Substances; D011401:Promoter Regions, Genetic; D011953:Receptors, Cyclic AMP / Q000502:physiology*; D011994:Recombinant Proteins / Q000378:metabolism; D013329:Structure-Activity Relationship; D014158:Transcription, Genetic
D046911:Macromolecular Substances; D011953:Receptors, Cyclic AMP; D011994:Recombinant Proteins; D000242:Cyclic AMP; D012321:DNA-Directed RNA Polymerases