Guanine nucleotides modulate the effects of brefeldin A in semipermeable cells: regulation of the association of a 110-kD peripheral membrane protein with the Golgi apparatus.
The release of a 110-kD peripheral membrane protein from the Golgi apparatus is an early event in brefeldin A (BFA) action, preceding the movement of Golgi membrane into the ER. ATP depletion also causes the reversible redistribution of the 110-kD protein from Golgi membrane into the cytosol, although no Golgi disassembly occurs. To further define the effects of BFA on the association of the 110-kD protein with the Golgi apparatus we have used filter perforation techniques to produce semipermeable cells. All previously observed effects of BFA, including the rapid redistribution of the 110-kD protein and the movement of Golgi membrane into the ER, could be reproduced in the semipermeable cells. The role of guanine nucleotides in this process was investigated using the nonhydrolyzable analogue of GTP, GTP gamma S. Pretreatment of semipermeable cells with GTP gamma S prevented the BFA-induced redistribution of the 110-kD protein from the Golgi apparatus and movement of Golgi membrane into the ER. GTP gamma S could also abrogate the observed release of the 110-kD protein from Golgi membranes which occurred in response to ATP depletion. Additionally, when the 110-kD protein had first been dissociated from Golgi membranes by ATP depletion, GTP gamma S could restore Golgi membrane association of the 110-kD protein, but not if BFA was present. All of these effects observed with GTP gamma S in semipermeable cells could be reproduced in intact cells treated with AlF4-. These results suggest that guanine nucleotides regulate the dynamic association/dissociation of the 110-kD protein with the Golgi apparatus and that BFA perturbs this process by interfering with the association of the 110-kD protein with the Golgi apparatus.
Donaldson
J G
JG
Cell Biology and Metabolism Branch, National Institute of Child Health and Human Development, National Institutes of Health, Bethesda, Maryland 20892.
Lippincott-Schwartz
J
J
Klausner
R D
RD
D000255:Adenosine Triphosphate / Q000502:physiology; D000535:Aluminum / Q000494:pharmacology; D017607:Aluminum Compounds; D000818:Animals; D020126:Brefeldin A; D002463:Cell Membrane Permeability; D002478:Cells, Cultured; D003517:Cyclopentanes / Q000494:pharmacology*; D003600:Cytosol / Q000378:metabolism; D005459:Fluorides; D005461:Fluorine / Q000494:pharmacology; D006056:Golgi Apparatus / Q000187:drug effects* / Q000378:metabolism; D006150:Guanine Nucleotides / Q000502:physiology*; D016244:Guanosine 5'-O-(3-Thiotriphosphate) / Q000494:pharmacology; D008565:Membrane Proteins / Q000378:metabolism*; D008970:Molecular Weight
D017607:Aluminum Compounds; D003517:Cyclopentanes; D006150:Guanine Nucleotides; D008565:Membrane Proteins; D020126:Brefeldin A; C050992:tetrafluoroaluminate; D005461:Fluorine; D016244:Guanosine 5'-O-(3-Thiotriphosphate); D000255:Adenosine Triphosphate; D000535:Aluminum; D005459:Fluorides