Nucleic Acids Research Advance Access originally published online on November 16, 2006
Nucleic Acids Research 2006 34(21):6362-6377; doi:10.1093/nar/gkl903
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Nucleic Acids Research, 2006, Vol. 34, No. 21 6362-6377
© 2006 The Author(s)
This is an Open Access article distributed under the terms of the Creative Commons Attribution Non-Commercial License (http://creativecommons.org/licenses/by-nc/2.0/uk/) which permits unrestricted non-commercial use, distribution, and reproduction in any medium, provided the original work is properly cited.
Molecular Biology |
The oligomeric Rep protein of Mungbean yellow mosaic India virus (MYMIV) is a likely replicative helicase
Plant Molecular Biology Group, International Centre for Genetic Engineering and Biotechnology Aruna Asaf Ali Marg, New Delhi-110 067, India
*To whom correspondence should be addressed. Tel: +91 11 26189358; Fax: +91 11 26162316; Email: sunilm{at}icgeb.res.in
Received July 31, 2006. Revised October 11, 2006. Accepted October 13, 2006.
Geminiviruses replicate by rolling circle mode of replication (RCR) and the viral Rep protein initiates RCR by the site-specific nicking at a conserved nonamer (TAATATT
AC) sequence. The mechanism of subsequent steps of the replication process, e.g. helicase activity to drive fork-elongation, etc. has largely remained obscure. Here we show that Rep of a geminivirus, namely, Mungbean yellow mosaic India virus (MYMIV), acts as a replicative helicase. The Rep-helicase, requiring 
6 nt space for its efficient activity, translocates in the 3'
5' direction, and the presence of forked junction in the substrate does not influence the activity to any great extent. Rep forms a large oligomeric complex and the helicase activity is dependent on the oligomeric conformation (
24mer). The role of Rep as a replicative helicase has been demonstrated through ex vivo studies in Saccharomyces cerevisiae and in planta analyses in Nicotiana tabacum. We also establish that such helicase activity is not confined to the MYMIV system alone, but is also true with at least two other begomoviruses, viz., Mungbean yellow mosaic virus (MYMV) and Indian cassava mosaic virus (ICMV).
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