Kinetic analysis of DNA and RNA strand transfer reactions catalyzed by vaccinia topoisomerase.

The Journal of Biological Chemistry
J SekiguchiS Shuman

Abstract

Vaccinia topoisomerase binds duplex DNA and forms a covalent DNA-(3'-phosphotyrosyl) protein adduct at the sequence 5'-CCCTT downward arrow. The enzyme reacts readily with a 36-mer CCCTT strand (DNA-p-RNA) composed of DNA 5' and RNA 3' of the scissile bond. However, a 36-mer composed of RNA 5' and DNA 3' of the scissile phosphate (RNA-p-DNA) is a poor substrate for covalent adduct formation. Vaccinia topoisomerase efficiently transfers covalently held CCCTT-containing DNA to 5'-OH-terminated RNA acceptors; the topoisomerase can therefore be used to tag the 5' end of RNA in vitro. Religation of the covalently bound CCCTT-containing DNA strand to a 5'-OH-terminated DNA acceptor is efficient and rapid (krel > 0.5 s-1), provided that the acceptor DNA is capable of base pairing to the noncleaved DNA strand of the topoisomerase-DNA donor complex. The rate of strand transfer to DNA is not detectably affected by base mismatches at the 5' nucleotide of the acceptor strand. Nucleotide deletions and insertions at the 5' end of the acceptor slow the rate of religation; the observed hierarchy of reaction rates is as follows: +1 insertion > -1 deletion > +2 insertion >> -2 deletion. These findings underscore the importance of a properly posi...Continue Reading

References

Nov 15, 1991·Proceedings of the National Academy of Sciences of the United States of America·S Shuman
Apr 20, 1986·Journal of Molecular Biology·S ArnottH S Park
Dec 1, 1989·Proceedings of the National Academy of Sciences of the United States of America·S ShumanS G Morham
Jan 23, 1996·Proceedings of the National Academy of Sciences of the United States of America·J SekiguchiS Shuman
Oct 15, 1996·Proceedings of the National Academy of Sciences of the United States of America·S W YangH A Nash
Feb 15, 1997·Nucleic Acids Research·J Sekiguchi, S Shuman

❮ Previous
Next ❯

Citations

Dec 5, 2002·Archives of Biochemistry and Biophysics·Hideyuki YamamotoEishichi Miyamoto
Sep 28, 1998·Biochimica Et Biophysica Acta·S Shuman
Apr 23, 2009·Molecules : a Journal of Synthetic Chemistry and Natural Product Chemistry·Agnes L-F ChanJau-Lang Hwang
Jul 11, 1998·Molecular Cell·J Sekiguchi, S Shuman
Jun 13, 1998·The Journal of Biological Chemistry·C Cheng, S Shuman
Jul 31, 1999·The Journal of Biological Chemistry·Y WangA H Andersen
Oct 3, 1998·The Journal of Biological Chemistry·T StraubF Boege
Jun 30, 2000·Journal of Molecular Biology·M KlemmA M Segall

❮ Previous
Next ❯

Related Concepts

Related Feeds

ASBMB Publications

The American Society for Biochemistry and Molecular Biology (ASBMB) includes the Journal of Biological Chemistry, Molecular & Cellular Proteomics, and the Journal of Lipid Research. Discover the latest research from ASBMB here.