TAILIEUCHUNG - Báo cáo khoa học: Minor capsid proteins of mouse polyomavirus are inducers of apoptosis when produced individually but are only moderate contributors to cell death during the late phase of viral infection

Minor structural proteins of mouse polyomavirus (MPyV) are essential for virus infection. To study their properties and possible contributions to cell death induction, fusion variants of these proteins, created by linking enhanced green fluorescent protein (EGFP) to their C- or N-termini, were prepared and tested in the absence of other MPyV gene products, namely the tumor antigens and the major capsid protein, VP1. | Minor capsid proteins of mouse polyomavirus are inducers of apoptosis when produced individually but are only moderate contributors to cell death during the late phase of viral infection Sandra Huerfano Vojtech Zila Evzen Boura Hana Spanielova Jitka Stokrova and Jitka Forstova Department of Genetics and Microbiology Faculty of Science Charles University Prague Czech Republic Keywords apoptosis minor proteins mouse polyomavirus VP2 VP3 Correspondence J. Forstova Department of Genetics and Microbiology Charles University in Prague ViniỉónỂỉ 5 128 44 Prague 2 Czech Republic Fax 420 2 21951729 Tel 420 2 21951730 E-mail jitkaf@ Received 4 November 2009 revised 15 December 2009 accepted 22 December 2009 doi Minor structural proteins of mouse polyomavirus MPyV are essential for virus infection. To study their properties and possible contributions to cell death induction fusion variants of these proteins created by linking enhanced green fluorescent protein EGFP to their C- or N-termini were prepared and tested in the absence of other MPyV gene products namely the tumor antigens and the major capsid protein VP1. The minor proteins linked to EGFP at their C-terminus VP2-EGFP VP3-EGFP were found to display properties similar to their nonfused wild-type versions they killed mouse 3T3 cells quickly when expressed individually. Carrying nuclear localization signals at their common C-terminus the minor capsid proteins were detected in the nucleus. However a substantial subpopulation of both VP2 and VP3 proteins as well as of the fusion proteins VP2-EGFP and VP3-EGFP was detected in the cytoplasm co-localizing with intracellular membranes. Truncated VP3 protein composed of 103 C-terminal amino acids exhibited reduced affinity for intracellular membranes and cytotoxicity. Biochemical studies proved each of the minor proteins to be a very potent inducer of apoptosis which was dependent on caspase activation. Immunoelectron microscopy .

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