Subcellular Localization of the Galphai3 Protein and G Alpha Interacting Protein, Two Proteins Involved in the Control of Macroautophagy in Human Colon Cancer HT-29 Cells

Overview

Journal Biochem J

Specialty Biochemistry

Date 1999 Jan 12

PMID 9882627

Citations 15

Authors

A Petiot

E Ogier-Denis

C Bauvy

F Cluzeaud

A Vandewalle

P Codogno

Affiliations

Soon will be listed here.

Abstract

Autophagic sequestration is controlled by the Galphai3 protein in human colon cancer HT-29 cells. Immunofluorescence and subcellular fractionation studies showed that the Galphai3 protein is preferentially associated with Golgi membranes but co-localization was also observed with the endoplasmic reticulum (ER) membrane. The Galphai2 protein, which is not involved in the control of autophagic sequestration, is associated with the plasma membrane. Transfection of chimaeric Galphai proteins (Galphai3/2, Galphai2/3) containing the N- and C-terminal parts of the relevant Galphai demonstrated that the C-terminal part of the Galphai3 protein, by governing its membrane localization [de Almeida, Holtzman, Peters, Ercolani, Ausiello and Stow (1994) J. Cell Sci. 107, 507-515], is important in the control of macroautophagic sequestration. G alpha interacting protein (GAIP),which stimulates the GTPase activity of the Galphai3 protein and favours macroautophagic sequestration in HT-29 cells,was shown, by immunofluorescence studies using confocal microscopy, to be confined to the cytoplasm. The cytoplasmic distribution of GAIP only partially overlaps with that of the Galphai3 protein. However, the presence of the two proteins on Golgi and ER membranes was confirmed by subcellular fractionation. These results point to the importance of the cytoplasmic localization of the Galphai3 protein and GAIP in controlling autophagic sequestration in HT-29 cells.

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References

Huang C, Hepler J, GILMAN A, Mumby S . Attenuation of Gi- and Gq-mediated signaling by expression of RGS4 or GAIP in mammalian cells. Proc Natl Acad Sci U S A. 1997; 94(12):6159-63. PMC: 21019. DOI: 10.1073/pnas.94.12.6159. View

Rabouille C, Strous G, Crapo J, Geuze H, Slot J . The differential degradation of two cytosolic proteins as a tool to monitor autophagy in hepatocytes by immunocytochemistry. J Cell Biol. 1993; 120(4):897-908. PMC: 2200086. DOI: 10.1083/jcb.120.4.897. View

Seglen P, Bohley P . Autophagy and other vacuolar protein degradation mechanisms. Experientia. 1992; 48(2):158-72. DOI: 10.1007/BF01923509. View

De Camilli P, Emr S, McPherson P, Novick P . Phosphoinositides as regulators in membrane traffic. Science. 1996; 271(5255):1533-9. DOI: 10.1126/science.271.5255.1533. View

Yamamoto A, Masaki R, Tashiro Y . Characterization of the isolation membranes and the limiting membranes of autophagosomes in rat hepatocytes by lectin cytochemistry. J Histochem Cytochem. 1990; 38(4):573-80. DOI: 10.1177/38.4.2319125. View

de Vries L, Mousli M, WURMSER A, Farquhar M . GAIP, a protein that specifically interacts with the trimeric G protein G alpha i3, is a member of a protein family with a highly conserved core domain. Proc Natl Acad Sci U S A. 1995; 92(25):11916-20. PMC: 40514. DOI: 10.1073/pnas.92.25.11916. View

Codogno P, Ogier-Denis E, Houri J . Signal transduction pathways in macroautophagy. Cell Signal. 1997; 9(2):125-30. DOI: 10.1016/s0898-6568(96)00130-1. View

Ogier-Denis E, Couvineau A, Maoret J, Houri J, Bauvy C, De Stefanis D . A heterotrimeric Gi3-protein controls autophagic sequestration in the human colon cancer cell line HT-29. J Biol Chem. 1995; 270(1):13-6. DOI: 10.1074/jbc.270.1.13. View

Stoyanov B, Volinia S, Hanck T, Rubio I, Loubtchenkov M, Malek D . Cloning and characterization of a G protein-activated human phosphoinositide-3 kinase. Science. 1995; 269(5224):690-3. DOI: 10.1126/science.7624799. View

10.

ZWEIBAUM A, Pinto M, Chevalier G, Dussaulx E, Triadou N, Lacroix B . Enterocytic differentiation of a subpopulation of the human colon tumor cell line HT-29 selected for growth in sugar-free medium and its inhibition by glucose. J Cell Physiol. 1985; 122(1):21-9. DOI: 10.1002/jcp.1041220105. View

11.

Dunn Jr W . Studies on the mechanisms of autophagy: formation of the autophagic vacuole. J Cell Biol. 1990; 110(6):1923-33. PMC: 2116114. DOI: 10.1083/jcb.110.6.1923. View

12.

Blommaart E, Luiken J, Meijer A . Autophagic proteolysis: control and specificity. Histochem J. 1997; 29(5):365-85. DOI: 10.1023/a:1026486801018. View

13.

Seglen P, Gordon P . 3-Methyladenine: specific inhibitor of autophagic/lysosomal protein degradation in isolated rat hepatocytes. Proc Natl Acad Sci U S A. 1982; 79(6):1889-92. PMC: 346086. DOI: 10.1073/pnas.79.6.1889. View

14.

Tooze J, Hollinshead M, Ludwig T, Howell K, Hoflack B, Kern H . In exocrine pancreas, the basolateral endocytic pathway converges with the autophagic pathway immediately after the early endosome. J Cell Biol. 1990; 111(2):329-45. PMC: 2116176. DOI: 10.1083/jcb.111.2.329. View

15.

Kadowaki M, Venerando R, Miotto G, MORTIMORE G . De novo autophagic vacuole formation in hepatocytes permeabilized by Staphylococcus aureus alpha-toxin. Inhibition by nonhydrolyzable GTP analogs. J Biol Chem. 1994; 269(5):3703-10. View

16.

Stromhaug P, Seglen P . Evidence for acidity of prelysosomal autophagic/endocytic vacuoles (amphisomes). Biochem J. 1993; 291 ( Pt 1):115-21. PMC: 1132489. DOI: 10.1042/bj2910115. View

17.

Lucocq J, Walker D . Evidence for fusion between multilamellar endosomes and autophagosomes in HeLa cells. Eur J Cell Biol. 1997; 72(4):307-13. View

18.

Lawrence B, Brown W . Autophagic vacuoles rapidly fuse with pre-existing lysosomes in cultured hepatocytes. J Cell Sci. 1992; 102 ( Pt 3):515-26. DOI: 10.1242/jcs.102.3.515. View

19.

Berman D, Wilkie T, GILMAN A . GAIP and RGS4 are GTPase-activating proteins for the Gi subfamily of G protein alpha subunits. Cell. 1996; 86(3):445-52. DOI: 10.1016/s0092-8674(00)80117-8. View

20.

Berman D, GILMAN A . Mammalian RGS proteins: barbarians at the gate. J Biol Chem. 1998; 273(3):1269-72. DOI: 10.1074/jbc.273.3.1269. View