1887

Abstract

The cucumber mosaic virus (CMV) replication complex has previously been shown to associate with cellular membranes. However, it remains unknown whether any host factors participate in this process. In this study, five groups of tonoplast intrinsic protein (TIP) genes were isolated and the proteins they encoded were evaluated with regard to their interactions with CMV proteins. TIP1 and TIP2 were found to interact with the CMV 1a protein in the Sos recruitment system, whereas no interactions with the other three TIP subgroups were observed in this assay. The interaction of CMV 1a with the TIP1 and TIP2 proteins was confirmed via co-immunoprecipitation assays. Additionally, CMV 1a co-localized with TIP1 and TIP2 in transfected protoplasts. The findings of this study suggest that members of two TIP subfamilies might affect CMV replication via interaction with CMV 1a in the tonoplasts.

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2006-11-01
2019-11-12
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vol. , part 11, pp. 3425-3431

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Interaction analyses of tonoplast intrinsic proteins (TIPs) and cucumber mosaic virus (CMV) or tobacco mosaiv virus (TMV) proteins using the yeast two-hybrid system. (a) Interaction analyses of TIPs and CMV 1a. cDNA encoding CMV 1a was fused with the activation domain in pGADT7, and different classes of cDNAs encoding TIPs were fused with the binding domain in pGBKT7. (b) Interaction analyses of TIPs and TMV replicase or helicase. cDNA encoding TMV replicase (nt 276-3419 of TMV; GenBank accession no. V01408) or helicase (nt 2553-3323) was fused with the activation domain. The two fusion vectors were co-transformed into the Y187 yeast strain, selected on plates devoid of Leu and Trp, and then subjected to colony-lift filter assays using X-Gal as a substrate. The pVA3-1/pTD1-1 and unfused pGADT7/pGBKT7 vectors were utilized as positive and negative controls, respectively. The blue spots indicate β-galactosidase activity.

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