Mono(ADP-ribosyl)ation of poly(ADP-ribose)polymerase by cholera toxin.

Poly(ADP-ribose)polymerase (PADPRP) was found to be an efficient protein acceptor for the arginine-specific ADP-ribosylation reaction catalyzed by cholera toxin (CT). The covalent modification of PADPRP was carried out with [32P]2'-dNAD as a selective mono(ADP-ribosyl)ation substrate. Mono(2'-dADP-ribosyl)ated-PADPRP was identified by autoradiographic analysis of the CT reaction products following sodium dodecyl sulfate-polyacrylamide gel electrophoresis. Addition of recombinant ADP-ribosylation factor (rARF), a small GTP-binding protein that stimulates the enzymatic activity of CT, enhanced the mono(2'-dADP-ribosyl)ation of PADPRP in a time- and substrate-dependent manner. In contrast, rARF did not change the ADP-ribose polymerizing activity of PADPRP. Peptide mapping mapping of [32P] labeled (2'-dADP-ribose)-PADPRP, following partial proteolysis with papain, revealed that the DNA-binding domain of PADPRP contained the mono(2'-dADP-ribosyl)ated arginine residue(s). Our results are consistent with the conclusion that PADPRP is susceptible to arginine-specific mono(ADP-ribosyl)ation catalyzed by CT.