Conformational change in the herpes simplex single-strand binding protein induced by DNA.
Protease digestion of the herpes simplex virus type 1 major single-strand DNA binding protein ICP8 showed that the cleavage patterns observed in the presence and absence of single-stranded DNA oligonucleotides are substantially different with protection of cleavage sites between amino acids 293 and 806 observed in the presence of oligonucleotide. Experiments using ICP8 modified with fluorescein-5-maleimide (FM) showed that the fluorescence signal exhibited increased susceptibility to antibody quenching and a significant decrease in polarization of the FM fluorescence was observed in the presence compared to the absence of oligonucleotide. Taken together, these results indicate that ICP8 undergoes a conformational change upon binding to single-stranded DNA.
Dudas
K C
KC
Department of Microbiology, Witebsky Center for Microbial Pathogenesis and Immunology, State University of New York at Buffalo, 3435 Main Street, Buffalo, NY 14214, USA.
Scouten
S K
SK
Ruyechan
W T
WT
D000906:Antibodies / Q000276:immunology; D004277:DNA, Single-Stranded / Q000378:metabolism*; D004268:DNA-Binding Proteins / Q000737:chemistry; D010450:Endopeptidases / Q000737:chemistry; D005452:Fluoresceins / Q000737:chemistry; D005456:Fluorescent Dyes / Q000737:chemistry; D009838:Oligodeoxyribonucleotides / Q000378:metabolism; D011487:Protein Conformation; D014764:Viral Proteins / Q000737:chemistry* / Q000378:metabolism
D000906:Antibodies; D004277:DNA, Single-Stranded; D004268:DNA-Binding Proteins; D005452:Fluoresceins; D005456:Fluorescent Dyes; C045462:ICP8 protein, Simplexvirus; D009838:Oligodeoxyribonucleotides; D014764:Viral Proteins; C037041:fluorescein 5-maleimide; D010450:Endopeptidases