PMID: 3754150Jan 14, 1986Paper

Identification of a nucleoside triphosphate binding site on calf thymus RNA polymerase II

Biochemistry
E Freund, P M McGuire

Abstract

A nucleoside triphosphate binding site on calf thymus RNA polymerase II was identified by using photoaffinity analogues of adenosine 5'-triphosphate and guanosine 5'-triphosphate. Both radiolabeled 8-azidoadenosine 5'-triphosphate (8-N3ATP) and radiolabeled 8-azidoguanosine 5'-triphosphate (8-N3GTP) bound to a single polypeptide of this enzyme. This polypeptide has a molecular mass of 37 kilodaltons and an isoelectric point of 5.4. Ultraviolet (UV) irradiation was necessary for photolabeling to occur. In addition, no labeling occurred when the probe was prephotolyzed or when the enzyme was inactivated. Furthermore, photolabeling of the enzyme could be decreased by preincubation with natural substrates. To provide evidence that the radiolabeled polypeptide forms a part of the domain of the nucleoside triphosphate binding site, experiments were performed using unlabeled 8-N3ATP. Although this unlabeled analogue was not a substrate for RNA polymerase II, it photoinactivated the enzyme in the presence of UV irradiation, and it inhibited transcription elongation by the enzyme in a competitive manner in the absence of UV irradiation. As in the case with photolabeling, photoinactivation by 8-N3ATP could be decreased by natural substra...Continue Reading

References

Feb 16, 1978·Biochimica Et Biophysica Acta·E G Kranias, R A Jungmann
Jan 1, 1979·Annual Review of Genetics·T Yura, A Ishihama
Jul 17, 1978·European Journal of Biochemistry·P ScheurichK Dose
Feb 16, 1978·Biochimica Et Biophysica Acta·E G Kranias, R A Jungmann
Mar 30, 1978·Biochemical and Biophysical Research Communications·P ValenzuelaJ Martial
Jan 1, 1977·Methods in Enzymology·H Bayley, J R Knowles
Jan 1, 1984·Annual Review of Biochemistry·P H von HippelJ A McSwiggen
Nov 18, 1983·Science·R A Young, R W Davis
Oct 1, 1983·Proceedings of the National Academy of Sciences of the United States of America·A S GordonI Diamond

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Citations

May 29, 1987·Biochemical and Biophysical Research Communications·R Y Chuang, L F Chuang

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