Stimulation of endogenous ADP-ribosylation by brefeldin A.
Brefeldin A (BFA) is a fungal metabolite that exerts profound and generally inhibitory actions on membrane transport. At least some of the BFA effects are due to inhibition of the GDP-GTP exchange on the ADP-ribosylation factor (ARF) catalyzed by membrane protein(s). ARF activation is likely to be a key event in the association of non-clathrin coat components, including ARF itself, onto transport organelles. ARF, in addition to participating in membrane transport, is known to function as a cofactor in the enzymatic activity of cholera toxin, a bacterial ADP-ribosyltransferase. In this study we have examined whether BFA, in addition to inhibiting membrane transport, might affect endogenous ADP-ribosylation in eukaryotic cells. Two cytosolic proteins of 38 and 50 kDa were enzymatically ADP-ribosylated in the presence of BFA in cellular extracts. The 38-kDa substrate was tentatively identified as the glycolytic enzyme glyceraldehyde-3-phosphate dehydrogenase. The BFA-binding components mediating inhibition of membrane traffic and stimulation of ADP-ribosylation appear to have the same ligand specificity. These data demonstrate the existence of a BFA-sensitive mono(ADP-ribosyl)transferase that may play a role in membrane movements.
De Matteis
M A
MA
Unit of Physiopathology of Secretion, Istituto di Ricerche Farmacologiche Mario Negri, Consorzio Mario Negri Sud, S. Maria Imbaro (Chieti), Italy.
Di Girolamo
M
M
Colanzi
A
A
Pallas
M
M
Di Tullio
G
G
McDonald
L J
LJ
Moss
J
J
Santini
G
G
Bannykh
S
S
Corda
D
D
D036002:ADP Ribose Transferases; D020727:ADP-Ribosylation Factors; D000246:Adenosine Diphosphate Ribose / Q000378:metabolism*; D000818:Animals; D001693:Biological Transport, Active; D020126:Brefeldin A; D002460:Cell Line; D003517:Cyclopentanes / Q000494:pharmacology*; D019204:GTP-Binding Proteins / Q000378:metabolism*; D005987:Glyceraldehyde-3-Phosphate Dehydrogenases / Q000378:metabolism; D009183:Mycotoxins / Q000494:pharmacology*; D009243:NAD / Q000378:metabolism; D011065:Poly(ADP-ribose) Polymerases / Q000378:metabolism; D011506:Proteins / Q000378:metabolism; D051381:Rats; D013379:Substrate Specificity
D003517:Cyclopentanes; D009183:Mycotoxins; D011506:Proteins; D009243:NAD; D020126:Brefeldin A; D000246:Adenosine Diphosphate Ribose; D005987:Glyceraldehyde-3-Phosphate Dehydrogenases; D036002:ADP Ribose Transferases; D011065:Poly(ADP-ribose) Polymerases; D019204:GTP-Binding Proteins; D020727:ADP-Ribosylation Factors