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J. Virol. doi:10.1128/JVI.01006-07
Copyright (c) 2007, American Society for Microbiology and/or the Listed Authors/Institutions. All Rights Reserved.

White spot syndrome virus proteins and differentially expressed host proteins identified in shrimp epithelium by shotgun proteomics and cleavable isotope-coded affinity tag

Department of Biological Sciences, National University of Singapore, Singapore 117543

^* To whom correspondence should be addressed. Email: dbshead{at}nus.edu.sg.

	Abstract

Shrimp subcuticular epithelial cells are the initial and major targets of white spot syndrome virus (WSSV) infection. Proteomic studies of WSSV-infected subcuticular epithelium of Peneaus monodon were performed through two approaches: subcellular fractionation coupled with shotgun proteomics to identify viral and host proteins, and a time-course quantitative proteomic analysis using cleavable isotope-coded affinity tags (cICAT) to identify differentially expressed cellular proteins. Peptides were analyzed by off-line coupling of two-dimensional liquid chromatography with MALDI-TOF/TOF mass spectrometry. We identified 27, 20 and 4 WSSV proteins from cytosolic, nuclear and membrane fractions, respectively. 28 unique WSSV proteins with high confidence (total ion C.I. %>95) were observed, of which 11 were reported here for the first time, and 3 of these novel proteins were shown to be viral non-structural proteins by western blotting analysis. A first shrimp protein dataset containing 1999 peptides (ion score 20) and 429 proteins (total ion score C.I. %>95) was constructed via shotgun proteomics. We also identified 10 down-regulated proteins and 2 up-regulated proteins from the shrimp epithelial lysate via cICAT analysis. This is the first comprehensive study of WSSV-infected epithelia by proteomics. These 11 novel viral proteins represent the latest addition to our knowledge of the WSSV proteome. Three proteomics datasets consisting of WSSV proteins, epithelial cellular proteins and differentially expressed cellular proteins generated in the course of WSSV infection provide a new resource to further study WSSV-shrimp interactions.