Poly(ADP-ribosyl)ation of a herpes simplex virus immediate early polypeptide.
This paper reports in vitro poly(ADP-ribosyl)ation of the herpes simplex virus type 1 (HSV-1) immediate early polypeptide Vmw175. The phenomenon was most clearly observed by use of the temperature-sensitive mutant tsK, which overproduces Vmw175 at the nonpermissive temperature (NPT) and has a mutation in the coding sequences for this polypeptide. Nuclei prepared from cells which were infected with tsK at NPT and subsequently downshifted to the permissive temperature incorporated [32P]NAD into Vmw175. This reaction did not occur when nuclei were prepared from cells constantly maintained at NPT, showing that only functional Vmw175 can be radiolabeled with [32P]NAD. The identity of the acceptor protein was confirmed by demonstrating the expected electrophoretic mobility differences between the HSV-1 and HSV-2 counterparts of Vmw175. The use of suitable inhibitors demonstrated that the reaction represented mono- or poly(ADP-ribosyl)ation, and further analysis showed the presence of long poly(ADP-ribose) chains attached to Vmw175. Poly(ADP-ribosyl)ation may be important as a cause or result of the regulation of viral transcription by Vmw175. Radiolabeling of another virus-specified polypeptide (approximate molecular weight 38,000), thought to be a structural component of the input virus, is also reported.
D004591:Electrophoresis, Polyacrylamide Gel; D009154:Mutation; D009243:NAD / Q000378:metabolism; D009702:Nucleoside Diphosphate Sugars / Q000378:metabolism*; D010455:Peptides / Q000378:metabolism*; D010761:Phosphorus Radioisotopes; D011064:Poly Adenosine Diphosphate Ribose / Q000032:analysis / Q000378:metabolism*; D018139:Simplexvirus / Q000378:metabolism*; D013696:Temperature; D014158:Transcription, Genetic; D014764:Viral Proteins / Q000378:metabolism*
D009702:Nucleoside Diphosphate Sugars; D010455:Peptides; D010761:Phosphorus Radioisotopes; D014764:Viral Proteins; D009243:NAD; D011064:Poly Adenosine Diphosphate Ribose