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.
Martinez
M
M
Department of Microbiology and Immunology, Texas College of Osteopathic Medicine/UNT, Fort Worth 76107-2690.
Price
S R
SR
Moss
J
J
Alvarez-Gonzalez
R
R
D036002:ADP Ribose Transferases; D000246:Adenosine Diphosphate Ribose / Q000378:metabolism*; D000818:Animals; D001345:Autoradiography; D002417:Cattle; D002772:Cholera Toxin / Q000494:pharmacology*; D004912:Erythrocytes / Q000201:enzymology; D007700:Kinetics; D009243:NAD / Q000031:analogs & derivatives / Q000138:chemical synthesis / Q000378:metabolism; D010761:Phosphorus Radioisotopes; D011065:Poly(ADP-ribose) Polymerases / Q000302:isolation & purification / Q000378:metabolism*; D013950:Thymus Gland / Q000201:enzymology; D014422:Turkeys
D010761:Phosphorus Radioisotopes; D009243:NAD; D000246:Adenosine Diphosphate Ribose; C021004:2'-deoxynicotinamide adenine dinucleotide; D002772:Cholera Toxin; D036002:ADP Ribose Transferases; D011065:Poly(ADP-ribose) Polymerases