Analysis of DNA-binding activity of the JC virus minor capsid protein VP2.
To investigate the DNA binding activity of the JC virus minor capsid protein, VP2, both wild-type and mutant VP2 were cloned and expressed in Escherichia coli. Southwestern blotting was employed for the DNA-binding assay. The results showed that VP2 was able to bind to DNA, except when either the last 13 or the last 29 amino acids were truncated. The results indicate that the DNA-binding domain of VP2 is located within the last 13 amino acids. Furthermore, we also demonstrated that Lys(332) and Lys(336) within the DNA-binding domain are crucial for DNA binding. The findings may provide further information for understanding the mechanism of virion assembly of the JC virus.
Huang
Yih-Leh
YL
Institute of Molecular Biology, National Chung Cheng University, Chia-Yi, Taiwan.
Wang
Meilin
M
Ou
Wei-Chih
WC
Fung
Chiung-Yau
CY
Chen
Li-Sheng
LS
Chang
Deching
D
D026501:Blotting, Southwestern; D036022:Capsid Proteins / Q000737:chemistry / Q000235:genetics* / Q000378:metabolism*; D003001:Cloning, Molecular; D004268:DNA-Binding Proteins / Q000235:genetics / Q000378:metabolism; D007577:JC Virus / Q000235:genetics*; D008239:Lysine / Q000235:genetics; D016296:Mutagenesis; D017354:Point Mutation; D017434:Protein Structure, Tertiary
D036022:Capsid Proteins; D004268:DNA-Binding Proteins; C442543:VP2 protein, Polyomavirus; D008239:Lysine