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A novel role for RAD54: this host protein modulates geminiviral DNA replication
Author(s) -
Kaliappan Kosalai,
Choudhury Nirupam Roy,
Suyal Geetika,
Mukherjee Sunil Kumar
Publication year - 2012
Publication title -
the faseb journal
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.709
H-Index - 277
eISSN - 1530-6860
pISSN - 0892-6638
DOI - 10.1096/fj.11-188508
Subject(s) - replication factor c , control of chromosome duplication , biology , dna replication , eukaryotic dna replication , dna replication factor cdt1 , origin recognition complex , ter protein , s phase , viral replication , replicon , microbiology and biotechnology , dna , virology , genetics , virus , plasmid
Geminiviruses primarily encode only few factors, such as replication initiator protein (Rep), and need various host cellular machineries for rolling‐circle replication (RCR) and/or recombination‐dependent replication (RDR). We have identified a host factor, RAD54, in a screen for Rep‐interacting partners and observed its role in DNA replication of the geminivirus mungbean yellow mosaic India virus (MYMIV). We identified the interacting domains Sc RAD54 and MYMIV‐Rep and observed that Sc RAD54 enhanced MYMIV‐Rep nicking, ATPase, and helicase activities. An in vitro replication assay demonstrated that the geminiviral DNA replication reaction depends on the viral Rep protein, viral origin of replication sequences, and host cell‐cycle proteins. Rad54‐deficient yeast nuclear extract did not support in vitro viral DNA replication, while exogenous addition of the purified Sc RAD54 protein enhanced replication. The role of RAD54 in in planta replication was confirmed by the transient replication assay; i.e. , agroinoculation studies. RAD54 is a well‐known recombination/repair protein that uses its DNA‐dependent ATPase activity in conjunction with several other host factors. However, this study demonstrates for the first time that the eukaryotic rolling‐circle replicon depends on the RAD54 protein.— Kaliappan, K., Choudhury, N. R., Suyal, G., Mukherjee, S. K. A novel role for RAD54: this host protein modulates geminiviral DNA replication. FASEB J. 26, 1142‐1160 (2012). www.fasebj.org