Regulated expression of the MADS-box transcription factor SrfA mediates activation of gene expression by protein kinase A during Dictyostelium sporulation.

Cell differentiation and morphogenesis are tightly regulated during sporulation in the lower eukaryote Dictyostelium discoideum. The control of the cyclic adenosine monophosphate (cAMP)-dependent protein kinase (PKA) is essential to coordinate these processes. Several signal transduction pathways are being recognized that lead to the regulation of intracellular cAMP levels. However, very little is known about the events lying downstream of PKA that are essential to activate late gene expression and terminal differentiation of the spores. We have studied the relationship between PKA and the MADS-box transcription factor SrfA, essential for spore differentiation. Constitutive activation of PKA was not able to rescue sporulation in a strain that lacks srfA suggesting the possibility that srfA functions downstream of PKA in a signal transduction pathway leading to spore maturation. A distal promoter region regulates the induction of srfA expression in the prespore region during culmination. We found that this promoter can be induced precociously by activating PKA with 8-Br-cAMP suggesting a transcriptional regulation by PKA. Moreover, precocious sporulation and expression of the spore marker spiA in a strain that overexpresses PKA, correlates with a precocious induction of srfA expression. The temporal and spatial pattern of expression was also studied in a mutant strain lacking the main adenylyl cyclase that functions during culmination, ACR. This strain is expected to have lower PKA activity and consistently, the level of srfA expression was reduced. Moreover, the temporal induction of srfA in the prespore region was also delayed during culmination. Our results strongly suggest that PKA activation during culmination leads to the induction of the expression of srfA. The correct temporal and spatial pattern of srfA expression appears to be part of a mechanism that ensures the adequate coordination of gene expression and morphogenesis. Copyright 2002 Elsevier Science Ireland Ltd.