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Journal of Virology, March 2004, p. 2758-2769, Vol. 78, No. 6
0022-538X/04/$08.00+0     DOI: 10.1128/JVI.78.6.2758-2769.2004
Copyright © 2004, American Society for Microbiology. All Rights Reserved.

Interaction between a Geminivirus Replication Protein and the Plant Sumoylation System

A. G. Castillo,1 L. J. Kong,2 L. Hanley-Bowdoin,2 and E. R. Bejarano1*

Department of Cellular Biology, Genetics and Animal Physiology, Málaga University, Málaga 29071, Spain,1 Department of Molecular and Structural Biochemistry, North Carolina State University, Raleigh, North Carolina 276952

Received 19 August 2003/ Accepted 14 November 2003

Geminiviruses are small DNA viruses that replicate in nuclei of infected plant cells after accumulation of host replication machinery. Tomato golden mosaic virus (TGMV) and Tomato yellow leaf curl Sardinia virus (TYLCSV) encode a protein, RepAC1 (or Rep), that is essential for viral replication. Rep/RepAC1 is an oligomeric protein that binds to double-stranded DNA, catalyzes cleavage and ligation of single-stranded DNA, and is sufficient for host induction. It also interacts with several host proteins, including the cell cycle regulator, retinoblastoma, and essential components of the cell DNA replication machinery, like proliferating nuclear cell antigen (PCNA) and RFC-1. To identify other cellular proteins that interact with Rep/RepAC1 protein, a Nicotiana benthamiana cDNA library was screened with a yeast two-hybrid assay. The host cell sumoylation enzyme, NbSCE1 (N. benthamiana SUMO-conjugating enzyme, homolog to Saccharomyces cerevisiae UBC9), was found to interact specifically with RepAC1. Mapping studies localized the interaction to the N-terminal half of RepAC1. Effects on geminivirus replication were observed in transgenic plants with altered levels of SUMO, the substrate for UBC9.

* Corresponding author. Mailing address: Department of Cellular Biology, Genetics and Animal Physiology, Málaga University, Málaga 29071, Spain. Phone: 34-952131677. Fax: 34-952131955. E-mail: edu_rodri{at}uma.es.

Journal of Virology, March 2004, p. 2758-2769, Vol. 78, No. 6
0022-538X/04/$08.00+0     DOI: 10.1128/JVI.78.6.2758-2769.2004
Copyright © 2004, American Society for Microbiology. All Rights Reserved.



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