Related Galpha subunits play opposing roles during Dictyostelium development

Of the several known Dictyostelium G protein subunits, the Galpha4 and Galpha5 subunits are the most closely related pair based on phylogenetic analysis and expression patterns, but these subunits perform different roles during development. To investigate potential relationships between these subunits with respect to cell differentiation, chimeric organisms composed of strains lacking or overexpressing either subunit were created and examined for developmental morphogenesis and spore production. Chimeras of galpha4 null and galpha5 null strains or Galpha4 and Galpha5 overexpression strains displayed compensatory morphogenesis, implying that the subunits promote complementary developmental processes. However, chimeras composed of galpha4 null and Galpha5 overexpression strains or galpha5 null and Galpha4 overexpression strains displayed distorted tip morphogenesis, suggesting the strains of these chimeras share common developmental deficiencies. Cells lacking the Galpha5 subunit localized to the prespore region of chimeras similar to the pattern observed for cells overexpressing the Galpha4 subunit, and cells overexpressing the Galpha5 subunit displayed localization patterns similar to galpha4 null mutants. A strain overexpressing both subunits displayed a partial suppression of morphology, gene expression, and cell localization phenotypes associated with the overexpression of the individual Galpha subunit genes, suggesting that each Galpha subunits can inhibit signaling mediated by the other subunit. Overexpression of the Galpha5 subunit inhibited chemotaxis and cGMP accumulation in response to folic acid, indicating that the Galpha5 subunit can inhibit early steps in the Galpha4-mediated signal transduction pathway. The contrasting phenotypes of the Galpha mutants suggest the Galpha4 and Galpha5 subunits provide opposing functions in cell differentiation, localization, and chemotactic responses to folic acid.