Initiation of (-)-strand RNA synthesis catalyzed by the BMV RNA-dependent RNA polymerase: synthesis of oligonucleotides

Virology
J H SunC C Kao

Abstract

RNA replication, a process of fundamental importance for pathogenesis by many viruses, remains poorly understood at the mechanistic level because relatively few of the responsible enzymes have been purified and characterized biochemically. Partially purified RNA-dependent RNA polymerase (RdRp) from brome mosaic virus (BMV)-infected barley leaves is able to synthesize (-)-strand RNAs from input (+)-strand templates. In resolving RdRp products generated during (-)-strand BMV RNA synthesis, we found that an RNA of eight nucleotides was generated at approximately 10-fold molar excess to the full-length (-)-strand RNA. Production of the 8-mer was dependent upon and specific to BMV RNA templates. Furthermore, inhibitors of full-length (-)-strand RNA synthesis by RdRp also affected the production of the 8-mer. Analysis of the sequence of the 8-mer suggests that it is initiated at the authentic initiation site on the BMV RNA template, consistent with the possibility that the 8-mer is an abortive initiation product of RNA synthesis by RdRp in vitro. Addition of heparin or Mn2+ differentially affected production of the 8-mer and full-length (-)-strand RNA, as did some nucleotide changes near the site of RNA initiation. These studies furt...Continue Reading

Citations

Mar 1, 2000·Molecular Plant Pathology·C C Kao, K Sivakumaran
Sep 3, 2004·Nucleic Acids Research·Jennifer T MillerStuart F J Le Grice
Oct 17, 2001·Journal of Virology·H YangD H Bamford
Apr 30, 2003·Journal of Virology·K SivakumaranC Cheng Kao
May 28, 2003·Journal of Virology·Jin-Hua SunMin Gao
Jan 15, 2003·Journal of Virology·C T Ranjith-KumarC Cheng Kao
Nov 10, 2001·Annual Review of Phytopathology·Theo W. Dreher
Oct 23, 1997·Proceedings of the National Academy of Sciences of the United States of America·R W SiegelC C Kao
Apr 20, 2001·Proceedings of the National Academy of Sciences of the United States of America·M J Kim, C Kao
Sep 30, 1998·Proceedings of the National Academy of Sciences of the United States of America·R W SiegelC C Kao
Aug 28, 2001·Nucleic Acids Research·R TayonC C Kao
Jan 31, 2004·Analytical Biochemistry·Frederick C Lahser, Bruce A Malcolm
Mar 11, 2004·Virus Research·Eugene V Makeyev, Jonathan M Grimes
Aug 15, 2015·Methods : a Companion to Methods in Enzymology·Radhakrishnan PadmanabhanKyung H Choi
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