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Interaction of Tomato Mosaic Virus Movement Protein with Tobacco RIO Kinase
Hiroshi Nyunoya,Kuniaki Yoshioka,Yasuhiko Matsushita,Masahiro Kasahara,Ken-ichi Konagaya 한국분자세포생물학회 2004 Molecules and cells Vol.17 No.2
Tomato mosaic virus (ToMV) has a regulatory gene encoding a movement protein (MP) that is involved in the cell-to-cell movement of viral RNA through plas-modesmata. To identify the host cell factors interacting with ToMV MP, we used a recombinant MP probe to isolate cDNA clones from a phage expression library of Nicotiana tabacum by a far-Western screening method. One of the cDNA clones encoded an MP-interacting protein, MIP-T7, homologous to the yeast novel pro-tein kinase, Rio1p. We isolated a full-length cDNA by RT-PCR. The putative gene product was designated NtRIO, and shared 33 and 73% amino acid identity with yeast and Arabidopsis RIO kinases, respectively. In vitro analyses using recombinant proteins showed that NtRIO also interacted with a different MP de-rived from Cucumber mosaic virus. NtRIO had auto-phosphorylation activity and phosphorylated ToMV MP. Addition of recombinant tobacco casein kinase 2 resulted in a marked increase in the phosphorylation of NtRIO. The interaction between NtRIO and ToMV MP was inhibited by phosphorylation of NtRIO.