Mapping of a contact for the RNA 3' terminus in the largest subunit of RNA polymerase.
Stalled elongation complexes of Escherichia coli RNA polymerase were prepared carrying the photo-cross-linkable 8-azido derivative of adenine at the 3'-terminus of the nascent RNA chain. Ultraviolet irradiation of such complexes resulted in the cross-linking of radiolabeled RNA exclusively to the beta' subunit of RNA polymerase. The adduct was mapped between Met932 and Trp1020 in the linear sequence of the beta' polypeptide using specific chemical degradation of the cross-linked species.
Borukhov
S
S
Department of Microbiology, Columbia University College of Physicians and Surgeons, New York, New York 10032.
Lee
J
J
Goldfarb
A
A
D000249:Adenosine Monophosphate / Q000031:analogs & derivatives / Q000378:metabolism; D000345:Affinity Labels / Q000378:metabolism; D000595:Amino Acid Sequence; D000818:Animals; D001386:Azides / Q000378:metabolism; D001665:Binding Sites; D012321:DNA-Directed RNA Polymerases / Q000235:genetics / Q000378:metabolism* / Q000528:radiation effects; D004926:Escherichia coli / Q000201:enzymology* / Q000235:genetics; D046911:Macromolecular Substances; D008969:Molecular Sequence Data; D012329:RNA, Bacterial / Q000235:genetics* / Q000378:metabolism; D012689:Sequence Homology, Nucleic Acid; D014158:Transcription, Genetic; D014466:Ultraviolet Rays
D000345:Affinity Labels; D001386:Azides; D046911:Macromolecular Substances; D012329:RNA, Bacterial; D000249:Adenosine Monophosphate; C021291:8-azidoadenosine 5'-monophosphate; D012321:DNA-Directed RNA Polymerases