J. Cell Biol., Volume 141, Number 7, June 29, 1998 1529-1537

G Protein  Subunit-null Mutants Are Impaired in Phagocytosis and Chemotaxis Due to Inappropriate Regulation of the Actin Cytoskeleton

Barbara Peracino,^* Jane Borleis, Tian Jin, Monika Westphal,^§ Jean-Marc Schwartz,^§ Lijun Wu, Enrico Bracco,^* Günther Gerisch,^§ Peter Devreotes, and Salvatore Bozzaro^*

^* Dipartimento di Scienze Cliniche e Biologiche, Università di Torino, Ospedale S. Luigi, 10043 Orbassano, Italy;  Department of Biological Chemistry, Johns Hopkins University School of Medicine, Baltimore, Maryland 21205; ^§ Max-Planck-Institut für Biochemie, 82152 Martinsried, Germany; and  Leukosite, Inc., Cambridge, Massachusetts 02142 

Chemotaxis and phagocytosis are basically similar in cells of the immune system and in Dictyostelium amebae. Deletion of the unique G protein  subunit in D. discoideum impaired phagocytosis but had little effect on fluid-phase endocytosis, cytokinesis, or random motility. Constitutive expression of wild-type  subunit restored phagocytosis and normal development. Chemoattractants released by cells or bacteria trigger typical transient actin polymerization responses in wild-type cells. In  subunit-null cells, and in a series of  subunit point mutants, these responses were impaired to a degree that correlated with the defect in phagocytosis. Image analysis of green fluorescent protein-actin transfected cells showed that  subunit- null cells were defective in reshaping the actin network into a phagocytic cup, and eventually a phagosome, in response to particle attachment. Our results indicate that signaling through heterotrimeric G proteins is required for regulating the actin cytoskeleton during phagocytic uptake, as previously shown for chemotaxis. Inhibitors of phospholipase C and intracellular Ca²⁺ mobilization inhibited phagocytosis, suggesting the possible involvement of these effectors in the process.

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