PROTEOMICS IN HOST VIRUS INTERACTION C.Anuradha and R.

Selvarajan Molecular Virology Lab, National Research Centre for Banana, Trichy, Tamil Nadu

Investigation of plant response to biotic and abiotic stress factors in the last decade revealed the existence of plant defence mechanisms of high complexity. Plant response to stress is no longer regarded as a linear succession of independent events, rather as a network of overlapping inter-regulated events, strongly influenced by plant growth and development as well as by environmental conditions. Cross-talk between the signalling pathways triggered by different stressors (eg., pathogen attack and herbivores/wounding) has been widely documented. Disentangling such complex defence systems require the investigation of plant reactions to different stressors in comparable conditions. Compatible plant-pathogen interactions are established when the plants fail to recognize the pathogen or when the pathogen suppresses the defence reaction of the host and result in plant susceptibility to the given pathogen. The hosts usually mobilize basic unspecific defence mechanisms and pathogen induced necrosis can trigger Systemic Acquired Resistance (SAR). Although patterns of response to pathogen attack common to many species have been described, the plants defence reactions against the pathogen are largely/solely host- and pathogen specific. Plant viruses exploit cellular factors, including host proteins, membranes and metabolites, for their replication in infected cells and to establish systemic infections. Proteomics constitute, nowadays, priority research for all host-pathogen interaction in the post-genomic era. The various proteomics approaches include measuring differential protein expression in virus-infected versus non-infected cells, analysis of viral and host protein components in the viral replicase or other virus-induced complexes, as well as proteome-wide screens to identify host protein - viral protein interactions using protein arrays or yeast twohybrid assays. A related concern is that substantial regulation of cellular events can occur at the protein level with no apparent changes in mRNA abundance. The posttranslational modification of proteins can result in a dramatic increase in protein complexity without a concomitant increase in gene expression. Proteomics may be the most promising technique for identification of proteins that are induced, repressed and post translationally modified during virus infection in plants.

The temporal patterns of protein expression during Rice Yellow Mottle Virus (RYMV) infection in rice cells of two cultivars: IR64, Oryza sativa indica, susceptible, and Azucena, O. sativa japonica, partially resistant to RYMV was studied through 2D-gel electrophoresis with mass spectrometry analysis. Proteomic analysis of non-stressed and RYMV inoculated cells showed statistically significant changes in the relative levels of 40 IR64 proteins and 24 Azucena proteins. Protein identification using mass spectrometry was attempted for all the differentially regulated proteins. This global analysis detected 32 hypothetical new proteins. Nineteen differentially regulated proteins were identified for IR64 cultivar, while 13 were identified for Azucena cultivar, including proteins in three functional categories: metabolism, stress-related proteins, and translation. These data revealed that a number of proteins regulated by abiotic stress response pathway were activated by RYMV in both cultivars (such as salt-induced protein, Heat Shock Proteins (HSPs), Superoxide Dismutase (SOD), and others which have functions consistent with susceptibility or the partially resistance trait (Ventelon-Debout et al., 2004). Proteomics-based approaches such as 2-DE and N-terminal sequencing were used to study the mechanisms responsible for the reduction of Oxygen Evolving Complex (OEC) protein content of Pepper Mild Mottle Virus-infected Nicotiana benthamiana plants (PerezBueno et al., 2004). A reduction in photosynthetic electron transport in photosystem II was investigated in virus-infected N. benthamiana plants using PAGE (Rahoutei et al., 2000) and this was the basis for performing a 2- DE analysis of changes in OEC polypeptide composition such as PsbP and PsbO. Perez-Bueno et al. (2004) observed a decrease in the amount of PsbP polypeptide of OEC as the disease progressed as compared to the PsbO polypeptide, and it was hypothesized that this difference might have a role in defense against viral infection. Differentially expressed proteins have been identified in soybean leaves infected with Soyabean Mosaic Virus (SMV) by proteomic approaches. This is the first extensive application of proteomics to the SMV-soybean interaction (Yang et al., 2011). In Capsicum chinense, as in other hosts, the infection by both compatible and incompatible viral strains of PMMoV induces the accumulation of a set of proteins. The 2-D approach allowed the identification of different Pathogenesis-Related (PR) protein isoforms in C. chinense leaves and their specific accumulation pattern in both the compatible and incompatible PMMoVC. chinense interactions, thus improving the results obtained by SDSPAGE analysis and allowing an easy identification of differentially expressed proteins in both type of interactions (Elvira et al., 2008).

Molecular events in host responses on viral diseases have not been studied intensively in plants till date. Hence, identification of differential expressed protein under normal and infected condition can reveal additional novel molecular pathways associated with disease development. While proteome-level investigations of plant-viral interactions has not been extensive, it is clear from the above studies that proteomics has the potential to contribute significantly in any future efforts to more comprehensively understand the interaction between plants and phytopathogenic viruses.