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Journal of Virology, November 1998, p. 9192-9200, Vol. 72, No. 11
0022-538X/98/$04.00+0
Copyright © 1998, American Society for Microbiology. All rights reserved.

Mutations in the N Terminus of the Brome Mosaic Virus Polymerase Affect Genetic RNA-RNA Recombination

M. Figlerowicz,1 P. D. Nagy,2 N. Tang,3 C. C. Kao,3 and J. J. Bujarski4,*

Institute of Bioorganic Chemistry, Polish Academy of Sciences, Poznan, Poland1; Department of Biochemistry and Molecular Biology, University of Massachusetts, Amherst, Massachusetts 010032; Department of Biology, Indiana University, Bloomington, Indiana 474053; and Plant Molecular Biology Center and Department of Biological Sciences, Northern Illinois University, De Kalb, Illinois 601154

Received 2 February 1998/Accepted 18 July 1998

Previously, we have observed that mutations in proteins 1a and 2a, the two virally encoded components of the brome mosaic virus (BMV) replicase, can affect the frequency of recombination and the locations of RNA recombination sites (P. D. Nagy, A. Dzianott, P. Ahlquist, and J. J. Bujarski, J. Virol. 69:2547-2556, 1995; M. Figlerowicz, P. D. Nagy, and J. J. Bujarski, Proc. Natl. Acad. Sci. USA 94:2073-2078, 1997). Also, it was found before that the N-terminal domain of 2a, the putative RNA polymerase protein, participates in the interactions between 1a and 2a (C. C. Kao, R. Quadt, R. P. Hershberger, and P. Ahlquist, J. Virol. 66:6322-6329, 1992; E. O'Reilly, J. Paul, and C. C. Kao, J. Virol. 71:7526-7532, 1997). In this work, we examine how mutations within the N terminus of 2a influence RNA recombination in BMV. Because of the likely electrostatic character of 1a-2a interactions, five 2a mutants, MF1 to MF5, were generated by replacing clusters of acidic amino acids with their neutral counterparts. MF2 and MF5 retained nearly wild-type levels of 1a-2a interaction and were infectious in Chenopodium quinoa. However, compared to that in wild-type virus, the frequency of nonhomologous recombination in both MF2 and MF5 was markedly decreased. Only in MF2 was the frequency of homologous recombination reduced and the occurrence of imprecise homologous recombination increased. In MF5 there was also a 3' shift in the positions of homologous crossovers. The observed effects of MF2 and MF5 reveal that the 2a N-terminal domain participates in different ways in homologous and in nonhomologous BMV RNA recombination. This work maps specific locations within the N terminus involved in 1a-2a interaction and in recombination and further suggests that the mechanisms of the two types of crossovers in BMV are different.

* Corresponding author. Mailing address: Plant Molecular Biology Center, Northern Illinois University, Montgomery Hall, De Kalb, IL 60115. Phone: (815) 753-0601. Fax: (815) 753-7810. E-mail: jbujarski{at}niu.edu.

Journal of Virology, November 1998, p. 9192-9200, Vol. 72, No. 11
0022-538X/98/$04.00+0
Copyright © 1998, American Society for Microbiology. All rights reserved.


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