title: Arabidopsis thaliana MCM2 plays role(s) in mungbean yellow mosaic India virus (MYMIV) DNA replication.
abstract: Geminiviruses are plant pathogens with single-stranded (ss) DNA genomes of about 2.7 kb in size. They replicate primarily via rolling-circle replication (RCR) with the help of a few virally encoded factors and various host-cell machineries. The virally encoded replication initiator protein (Rep) is essential for geminivirus replication. In this study, by interaction screening of an Arabidopsis thaliana cDNA library, we have identified a host factor, MCM2, that interacts with the Rep protein of the geminivirus mungbean yellow mosaic India virus (MYMIV). Using yeast two-hybrid, β-galactosidase and co-immunoprecipitation assays, we demonstrated an interaction between MYMIV-Rep and the host factor AtMCM2. We investigated the possible role of AtMCM2 in geminiviral replication using a yeast-based geminivirus DNA replication restoration assay and observed that the AtMCM2 protein complemented the mcm2∆ mutation of S. cerevisiae. Our data suggest the involvement of AtMCM2 in the replication of MYMIV ex vivo. The role of MCM2 in replication was confirmed in planta by a transient replication assay in both wild-type and mutant Arabidopsis plants through agroinoculation. Our data provide evidence for the involvement of AtMCM2 in geminiviral DNA replication, presumably in conjunction with other host factors, and suggest its importance in MYMIV DNA replication.
authors.affiliation: Plant Molecular Biology Group, International Centre for Genetic Engineering and Biotechnology ICGEB, Aruna Asaf Ali Marg, New Delhi 110 067, India. gits27@gmail.com
authors.forename: Geetika
authors.initials: G
authors.lastname: Suyal
authors.forename: Sunil K
authors.initials: SK
authors.lastname: Mukherjee
authors.forename: Prem S
authors.initials: PS
authors.lastname: Srivastava
authors.forename: Nirupam R
authors.initials: NR
authors.lastname: Choudhury
mesh_terms: D017360:Arabidopsis / Q000821:virology*; D029681:Arabidopsis Proteins / Q000235:genetics / Q000378:metabolism*; D053839:Begomovirus / Q000472:pathogenicity / Q000502:physiology*; D004265:DNA Helicases / Q000378:metabolism*; D004261:DNA Replication; D017353:Gene Deletion; D005816:Genetic Complementation Test; D054884:Host-Pathogen Interactions; D047468:Immunoprecipitation; D025941:Protein Interaction Mapping; D012441:Saccharomyces cerevisiae / Q000201:enzymology; D015534:Trans-Activators / Q000378:metabolism*; D020798:Two-Hybrid System Techniques; D001616:beta-Galactosidase / Q000032:analysis
chemical_list: D029681:Arabidopsis Proteins; D015534:Trans-Activators; C045264:replication initiator protein; D001616:beta-Galactosidase; D004265:DNA Helicases