The opportunistic human pathogen Penicillium marneffei exhibits a temperature-dependent dimorphic switch. At 25 degrees, multinucleate, septate hyphae that can undergo differentiation to produce asexual spores (conidia) are produced. At 37 degrees hyphae undergo arthroconidiation to produce uninucleate yeast cells that divide by fission. This work describes the cloning of the P. marneffei gasC gene encoding a G-protein alpha-subunit that shows high homology to members of the class III fungal Galpha-subunits. Characterization of a DeltagasC mutant and strains carrying a dominant-activating gasC(G45R) or a dominant-interfering gasC(G207R) allele show that GasC is a crucial regulator of germination. A DeltagasC mutant is severely delayed in germination, whereas strains carrying a dominant-activating gasC(G45R) allele show a significantly accelerated germination rate. Additionally, GasC signaling positively affects the production of the red pigment by P. marneffei at 25 degrees and negatively affects the onset of conidiation and the conidial yield, showing that GasC function overlaps with functions of the previously described Galpha-subunit GasA. In contrast to the S. cerevisiae ortholog Gpa2, our data indicate that GasC is not involved in carbon or nitrogen source sensing and plays no major role in either hyphal or yeast growth or in the switch between these two forms.