Differences in the poly(ADP-ribosyl)ation patterns of ICP4, the herpes simplex virus major regulatory protein, in infected cells and in isolated nuclei.
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.
Blaho
J A
JA
Marjorie B. Kovler Viral Oncology Laboratories, University of Chicago, Illinois 60637.
Michael
N
N
Kang
V
V
Aboul-Ela
N
N
Smulson
M E
ME
Jacobson
M K
MK
Roizman
B
B
D000249:Adenosine Monophosphate / Q000378:metabolism; D002461:Cell Line, Transformed; D002467:Cell Nucleus / Q000737:chemistry; D006026:Glycoside Hydrolases / Q000494:pharmacology; D006157:Guanosine Monophosphate / Q000378:metabolism; D006561:Herpes Simplex / Q000378:metabolism*; D006801:Humans; D017874:Immediate-Early Proteins; D009243:NAD / Q000378:metabolism; D009687:Nuclear Proteins / Q000187:drug effects; D011064:Poly Adenosine Diphosphate Ribose / Q000032:analysis / Q000276:immunology / Q000378:metabolism*; D011489:Protein Denaturation; D011499:Protein Processing, Post-Translational; D013379:Substrate Specificity; D054334:Viral Regulatory and Accessory Proteins / Q000737:chemistry / Q000302:isolation & purification / Q000378:metabolism*
D017874:Immediate-Early Proteins; D009687:Nuclear Proteins; D054334:Viral Regulatory and Accessory Proteins; C048925:herpes simplex virus, type 1 protein ICP4; D009243:NAD; D011064:Poly Adenosine Diphosphate Ribose; D000249:Adenosine Monophosphate; D006157:Guanosine Monophosphate; D006026:Glycoside Hydrolases; C022407:poly ADP-ribose glycohydrolase