Pseudorabies virus DNA-binding protein stimulates the exonuclease activity and regulates the processivity of pseudorabies virus DNase.
The pseudorabies virus (PRV) DNase is an alkaline exonuclease and endonuclease, which exhibits an Escherichia coli RecBCD-like catalytic function. The PRV DNA-binding protein (DBP) promotes the renaturation of complementary single strands of DNA, which is an essential function for recombinase. To investigate the functional and physical interactions between PRV DBP and DNase, these proteins were purified to homogeneity. PRV DBP stimulated the DNase activity, especially the exonuclease activity, in a dose-dependent fashion. Acetylation of DBP by acetic anhydride resulted in a loss of DNA-binding ability and a 60% inhibition of the DNase activity, suggesting that DNA-binding ability of PRV DBP was required for stimulating the DNase activity. PRV DNase behaved in a processive mode; however, it was converted into a distributive mode in the presence of DBP, implying that PRV DBP stimulated the dissociation of DNase from DNA substrates. The physical interaction between DBP and DNase was further analyzed by enzyme-linked immunosorbent assay, and a significant interaction was observed. Thus, these results suggested that PRV DBP interacted with PRV DNase and regulated the DNase activity in vitro.
Hsiang
Chien-Yun
CY
Department of Microbiology, China Medical College, 91 Hsueh-Shih Road, Taichung 404, Taiwan. cyhsiang@mail.cmc.edu.tw
D000089:Acetic Anhydrides / Q000494:pharmacology; D020134:Catalytic Domain; D003101:Collodion / Q000378:metabolism; D004247:DNA / Q000378:metabolism*; D003851:Deoxyribonucleases / Q000378:metabolism*; D004305:Dose-Response Relationship, Drug; D004797:Enzyme-Linked Immunosorbent Assay; D004926:Escherichia coli / Q000378:metabolism; D005092:Exonucleases / Q000378:metabolism*; D011558:Herpesvirus 1, Suid / Q000378:metabolism*; D008239:Lysine / Q000737:chemistry; D011485:Protein Binding; D011995:Recombination, Genetic; D013997:Time Factors; D014764:Viral Proteins / Q000378:metabolism*
D000089:Acetic Anhydrides; D014764:Viral Proteins; C031800:acetic anhydride; D003101:Collodion; D004247:DNA; D003851:Deoxyribonucleases; D005092:Exonucleases; D008239:Lysine