A newly identified interaction between IVa2 and pVIII proteins during porcine adenovirus type 3 infection

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• 作者：Singh ; Mahavir ; Shmulevitz ; Maya ; Tikoo ; Suresh Kumar
• 关键词：IVa2 ; pVIII ; Protein– ; protein interaction ; Yeast two-hybrid ; GST pull-down assay ; Co-immunoprecipitation ; Metabolic-labeling ; Confocal microscopy ; Viral assembly
• 刊名：Virology
• 出版年：2005
• 期刊代码：108_00426822
• 类别：imm
• 出版时间：May 25, 2005
• 卷：336
• 期：1
• 页码：60-69
• 文件大小：380 K

摘要

The adenovirus IVa2 is an intermediate viral gene product that appears to perform multiple essential roles in viral infection. Using IVa2 as bait in the yeast two-hybrid system, we screened selected open reading frames (ORFs) of porcine adenovirus (PAdV)-3 for potential interaction with IVa2. Interestingly, pVIII showed specific interaction with IVa2. The yeast two-hybrid findings were validated by GST pull-down assays, in vitro binding studies employing cell-free coupled transcription–translation products and in vitro co-immunoprecipitations using protein-specific antibodies. Finally, we demonstrated that IVa2 specifically interacts with pVIII during PAdV-3 infection.