Role of NAD(+) and ADP-ribosylation in the maintenance of the Golgi structure

被引：44

作者：

Mironov, A ^{[1
]}

Colanzi, A ^{[1
]}

Silletta, MG ^{[1
]}

Fiucci, G ^{[1
]}

Flati, S ^{[1
]}

Fusella, A ^{[1
]}

Polishchuk, R ^{[1
]}

Mironov, A ^{[1
]}

DiTullio, G ^{[1
]}

Weigert, R ^{[1
]}

Malhotra, V ^{[1
]}

Corda, D ^{[1
]}

DeMatteis, MA ^{[1
]}

Luini, A ^{[1
]}

机构：

[1] UNIV CALIF SAN DIEGO,DEPT BIOL,LA JOLLA,CA 92093

来源：

JOURNAL OF CELL BIOLOGY | 1997年 / 139卷 / 05期

关键词：

D O I：

10.1083/jcb.139.5.1109

中图分类号：

Q2 [细胞生物学];

学科分类号：

071009 ; 090102 ;

摘要：

We have investigated the role of the ADP-ribosylation induced by brefeldin A (BFA) in the mechanisms controlling the architecture of the Golgi complex. BFA causes the rapid disassembly of this organelle into a network of tubules, prevents the association of coatomer and other proteins to Golgi membranes, and stimulates the ADP-ribosylation of two cytosolic proteins of 38 and 50 kD (GAPDH and BARS-50; De Matteis, M.A., M. DiGirolamo, A. Colanzi, M. Pallas, G. Di Tullio, L.J. McDonald, J. Moss, G. Santini, S. Bannykh, D. Corda, and A. Luini. 1994. Proc. Natl. Acad Sci. USA. 91:1114-1118; Di Girolamo, M., M.G. Silletta, M.A. De Matteis, A. Braca, A. Colanzi, D. Pawlak, M.M. Rasenick, A. Luini, and D. Corda. 1995. Proc. Natl. Acad Sci USA. 92:7065-7069). To study the role of ADP-ribosylation, this reaction was inhibited by depletion of NAD(+) (the ADP-ribose donor) or by using selective pharmacological blockers in permeabilized cells. In NAD(+)-depleted cells and in the presence of dialized cytosol, BFA detached coat proteins from Golgi membranes with normal potency but failed to alter the organelle's structure. Readdition of NAD(+) triggered Golgi disassembly by BFA. This effect of NAD(+) was mimicked by the use of pre-ADP-ribosylated cytosol, The further addition of extracts enriched in native BARS-50 abolished the ability of ADP-ribosylated cytosol to support the effect of BFA. Pharmacological blockers of the BFA-dependent ADP-ribosylation (Weigert, R., A. Colanzi, A. Mironov, R. Buccione, C. Cericola, M.G. Sciulli, G. Santini, S. Flati, A. Fusella, J. Donaldson, M. DiGirolamo, D. Corda, M.A. De Matteis, and A. Luini. 1997. J. Biol. Chern. 272:14200-14207) prevented Golgi disassembly by BFA in permeabilized cells. These inhibitors became inactive in the presence of pre-ADP-ribosylated cytosol, and their activity was rescued by supple menting the cytosol with a native BARS50-enriched fraction. These results indicate that ADP-ribosylation plays a role in the Golgi disassembling activity of BFA, and suggest that the ADP-ribosylated substrates are components of the machinery controlling the structure of the Golgi apparatus.

引用

页码：1109 / 1118

页数：10

共 63 条

[1] THE FORMATION OF GOLGI STACKS FROM VESICULATED GOLGI MEMBRANES REQUIRES 2 DISTINCT FUSION EVENTS [J].