Interaction of Bombyx mori nucleopolyhedrovirus BRO-A and host cell protein laminin

• Published in: Archives of virology 2003, Vol.148 (1), p.99-113
• Main Authors: Kang, W. K, Imai, N, Suzuki, M, Iwanaga, M, Matsumoto, S, Zemskov, E. A
• Format: Article
• Language: English
• Subjects: 3' Flanking Region; 3' Untranslated Regions; Amino Acid Sequence; Animals; antibodies; Base Sequence; binding proteins; Biological and medical sciences; Bombyx; Bombyx - virology; Bombyx mori nucleopolyhedrovirus; cDNA libraries; clones; Cloning; complementary DNA; DNA-Binding Proteins - genetics; DNA-Binding Proteins - metabolism; Drosophila; Fundamental and applied biological sciences. Psychology; Genes; Genomes; Infections; Insects; laminin; Laminin - genetics; Laminin - metabolism; Microbiology; Molecular Sequence Data; Nucleopolyhedrovirus - genetics; Nucleopolyhedrovirus - metabolism; Nucleoproteins - metabolism; nucleosomes; Open Reading Frames; Plasmids; Proteins; Replicative cycle, interference, host-virus relations, pathogenicity, miscellaneous strains; Sequence Homology, Amino Acid; two hybrid system techniques; Virology
• ISSN: 0304-8608; 1432-8798
• DOI: 10.1007/s00705-002-0902-7

The Bombyx mori nucleopolyhedrovirus (BmNPV) contains five baculovirus repeated ORF (bro) genes, all of which are expressed as delayed early genes. We have recently reported that BmNPV BRO proteins, specially BRO-A and BRO-C, contain a nucleic acid binding activity and are involved in nucleosome structures in nuclei of infected cells. To further understand the function of bro-a gene, we looked for factors interacting with BmNPV BRO-A using the yeast two-hybrid system. Fifteen clones obtained from a cDNA library of mock-infected cells and one from a library prepared at 2 h postinfection (p.i.) were found to comprise one distinct gene, which was identified as the Bombyx homolog (bLaminin) of Drosophila laminin β1. A direct interaction between BRO-A and N-terminal region of bLaminin was demonstrated by in vitro pull-down experiments. Further pull-down assays using BmN cell extracts and anti-laminin antibodies also showed interaction of both proteins. In addition, two more clones were obtained from cDNA library of 12 h p.i. and were found to encode BRO-A itself, indicating that BRO-A forms an oligomer. Taken together, we propose that BRO-A may function as a laminin binding protein.