guanosine-monophosphate and Herpes-Simplex

Infected-cell protein 4 (ICP4), the major regulatory protein in herpes simplex viruses 1 and 2, was previously reported to accept 32P from [32P]NAD in isolated nuclei. This modification was attributed to poly(ADP-ribosyl)ation (C. M. Preston and E. L. Notarianni, Virology 131:492-501, 1983). We determined that an antibody specific for poly(ADP-ribose) reacts with ICP4 extracted from infected cells, electrophoretically separated in denaturing gels, and electrically transferred to nitrocellulose. Our results indicate that all forms of ICP4 observed in one-dimensional gel electrophoresis are poly(ADP-ribosyl)ated. Poly(ADP-ribose) on ICP4 extracted from infected cells was resistant to cleavage by purified poly(ADP-ribose) glycohydrolase unless ICP4 was in a denatured state. Poly(ADP-ribose) added to ICP4 in isolated nuclei was sensitive to this enzyme. This result indicates that the two processes are distinct and may involve different sites on the ICP4 molecule.

Topics: Adenosine Monophosphate; Cell Line, Transformed; Cell Nucleus; Glycoside Hydrolases; Guanosine Monophosphate; Herpes Simplex; Humans; Immediate-Early Proteins; NAD; Nuclear Proteins; Poly Adenosine Diphosphate Ribose; Protein Denaturation; Protein Processing, Post-Translational; Substrate Specificity; Viral Regulatory and Accessory Proteins