PMID: 7538173Jun 1, 1995Paper

A maltose-binding protein/adeno-associated virus Rep68 fusion protein has DNA-RNA helicase and ATPase activities

Journal of Virology
R S WonderlingR A Owens

Abstract

The adeno-associated virus type 2 (AAV) Rep68 protein produced in Escherichia coli as a fusion protein with maltose-binding protein (MBP-Rep68 delta) has previously been shown to possess DNA-DNA helicase activity, as does the purified wild-type Rep68. In the present study, we demonstrate that MBP-Rep68 delta also catalyzes the unwinding of a DNA-RNA hybrid. MBP-Rep68 delta-mediated DNA-RNA helicase activity required ATP hydrolysis and the presence of Mg2+ ions and was inhibited by high ionic strength. The efficiency of the DNA-RNA helicase activity of MBP-Rep68 delta was comparable to its DNA-DNA helicase activity. However, MBP-Rep68 delta lacked the ability to unwind a blunt-ended DNA-RNA substrate and RNA-RNA duplexes. We have also demonstrated that MBP-Rep68 delta has ATPase activity which is enhanced by the presence of single-stranded DNA but not by RNA. The MBP-Rep68 delta NTP mutant protein, which has a lysine-to-histidine substitution at amino acid 340 in the putative nucleoside triphosphate-binding site of Rep68, not only lacks DNA-RNA helicase and ATPase activities but also inhibits the helicase activity of MBP-Rep68 delta. DNA-RNA helicase activity of Rep proteins might play a pivotal role in the regulation of AAV gen...Continue Reading

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Citations

Jul 23, 2002·Journal of Virology·Daniel F Lackner, Nicholas Muzyczka
Jun 26, 2003·The Journal of Biological Chemistry·Roy F CollacoJames P Trempe
May 28, 2003·Journal of Virology·Ioannis Bossis, John A Chiorini
Sep 15, 2005·Proceedings of the National Academy of Sciences of the United States of America·Carole BerthetPeter Beard
Aug 17, 2011·Proceedings of the National Academy of Sciences of the United States of America·Varsha SitaramanWadie F Bahou
Sep 18, 2009·Journal of Virology·Nathalie DutheilR Michael Linden
May 26, 1999·The Plant Journal : for Cell and Molecular Biology·F L EricksonB Baker
Jan 9, 1999·Journal of Virology·J A ChioriniR M Kotin
Mar 30, 2017·Human Gene Therapy·Robert M Kotin, Richard O Snyder
May 4, 2007·Biochemistry·Susan S DignamJohn David Dignam
Dec 21, 2007·Biochemistry·Sarmistha BandyopadhyayPaul L Hermonat
Mar 3, 1997·Virology·K M PrasadJ P Trempe
Mar 1, 1997·Journal of Virology·R S Wonderling, R A Owens

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