Utilization of variably spaced promoter-like elements by the bacterial RNA polymerase holoenzyme during early elongation.
The bacterial RNA polymeras holoenzyme consists of a catalytic core enzyme in complex with a sigma factor that is required for promoter-specific transcription initiation. During initiation, members of the sigma(70) family of sigma factors contact two conserved promoter elements, the -10 and -35 elements, which are separated by approximately 17 base pairs (bp). sigma(70) family members contain four flexibly linked domains. Two of these domains, sigma(2) and sigma(4), contain determinants for interactions with the promoter -10 and -35 elements respectively. sigma(2) and sigma(4) also contain core-binding determinants. When bound to core the inter-domain distance between sigma(2) and sigma(4) matches the distance between promoter elements separated by approximately 17 bp. Prior work indicates that during early elongation the nascent RNA-assisted displacement of sigma(4) from core can enable the holoenzyme to adopt a configuration in which sigma(2) and sigma(4) are bound to 'promoter-like' DNA elements separated by a single base pair. Here we demonstrate that holoenzyme can also adopt configurations in which sigma(2) and sigma(4) are bound to 'promoter-like' DNA elements separated by 0, 2 or 3 bp. Thus, our findings suggest that displacement of sigma(4) from core enables the RNA polymerase holoenzyme to adopt a broad range of 'elongation-specific' configurations.
Devi
Pukhrambam Grihanjali
PG
Department of Genetics and Waksman Institute, Rutgers University, Piscataway, NJ 08854, USA.
Campbell
Elizabeth A
EA
Darst
Seth A
SA
Nickels
Bryce E
BE
D001419:Bacteria / Q000201:enzymology*; D012321:DNA-Directed RNA Polymerases / Q000737:chemistry / Q000378:metabolism*; D020035:Holoenzymes / Q000737:chemistry / Q000378:metabolism; D011401:Promoter Regions, Genetic; D012329:RNA, Bacterial / Q000378:metabolism; D012045:Regulatory Sequences, Nucleic Acid; D012808:Sigma Factor / Q000737:chemistry / Q000378:metabolism*; D014158:Transcription, Genetic; D014764:Viral Proteins / Q000378:metabolism
D020035:Holoenzymes; C066119:Q protein, Bacteriophage lambda; D012329:RNA, Bacterial; D012808:Sigma Factor; D014764:Viral Proteins; C081775:RNA polymerase sigma 70; D012321:DNA-Directed RNA Polymerases