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Vol. 12, Issue 10, 3204-3213, October 2001
in Dictyostelium: Its Role in Localization of
G
to the Membrane Is Required for Chemotaxis in Shallow Gradients
Department of Cell Biology and Anatomy, Johns Hopkins University
School of Medicine, Baltimore, Maryland 21205
G-protein-mediated signal transduction pathways play an essential
role in the developmental program of the simple eukaryotic organism
Dictyostelium discoideum. Database searches have yielded 11 G
-subunits, a single G
-subunit, but no G
-subunits. We
report here the purification, cDNA isolation, and functional analysis of a G-subunit. Like G, the G appears to be unique and
hybridization studies show that G and G are expressed in parallel
during development. Species-wide sequence comparisons of G-subunits
and -like domains of RGS proteins reveal short stretches of highly
conserved residues as well as the common CXXL motif at the
COOH-terminal of Gs that target Gs to plasma membrane.
Overexpression of a CSVL-deleted G (G
) in wild-type cells
shifts G to the cytosol and selectively impairs certain
G-protein-mediated signal transduction pathways. These cells are able
to respond to increments in the stimulus, but are unable to sense
chemoattractant gradients. They neither move directionally nor recruit
PH-domains to their leading edge. Thus, a full complement of
membrane-tethered G is required for sensing shallow gradients,
but is not essential for responses to increments in extracellular stimuli.
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