G protein beta subunit-null mutants are impaired in phagocytosis and chemotaxis due to inappropriate regulation of the actin cytoskeleton.
Chemotaxis and phagocytosis are basically similar in cells of the immune system and in Dictyostelium amebae. Deletion of the unique G protein beta subunit in D. discoideum impaired phagocytosis but had little effect on fluid-phase endocytosis, cytokinesis, or random motility. Constitutive expression of wild-type beta subunit restored phagocytosis and normal development. Chemoattractants released by cells or bacteria trigger typical transient actin polymerization responses in wild-type cells. In beta subunit-null cells, and in a series of beta 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 beta 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.
Peracino
B
B
Dipartimento di Scienze Cliniche e Biologiche, Università di Torino, Ospedale S. Luigi, 10043 Orbassano, Italy.
Borleis
J
J
Jin
T
T
Westphal
M
M
Schwartz
J M
JM
Wu
L
L
Bracco
E
E
Gerisch
G
G
Devreotes
P
P
Bozzaro
S
S
D000199:Actins / Q000502:physiology*; D000818:Animals; D002455:Cell Division; D002465:Cell Movement; D002630:Chemotactic Factors / Q000494:pharmacology; D002633:Chemotaxis / Q000502:physiology*; D003599:Cytoskeleton / Q000502:physiology; D004023:Dictyostelium / Q000378:metabolism / Q000502:physiology; D004926:Escherichia coli / Q000378:metabolism; D019204:GTP-Binding Proteins / Q000235:genetics / Q000502:physiology*; D016296:Mutagenesis; D010587:Phagocytosis / Q000502:physiology*; D010873:Pinocytosis; D012475:Salmonella / Q000378:metabolism; D015398:Signal Transduction
D000199:Actins; D002630:Chemotactic Factors; D019204:GTP-Binding Proteins