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Eukaryotic Cell, August 2006, p. 1287-1300, Vol. 5, No. 8
1535-9778/06/$08.00+0     doi:10.1128/EC.00109-06
Copyright © 2006, American Society for Microbiology. All Rights Reserved.

GPR-4 Is a Predicted G-Protein-Coupled Receptor Required for Carbon Source-Dependent Asexual Growth and Development in Neurospora crassa

Liande Li and Katherine A. Borkovich^*

Department of Plant Pathology, 1415 Boyce Hall, 900 University Avenue, University of California, Riverside, Riverside, California 92521

Received 15 April 2006/ Accepted 30 May 2006

The filamentous fungus Neurospora crassa is able to utilize a wide variety of carbon sources. Here, we examine the involvement of a predicted G-protein-coupled receptor (GPCR), GPR-4, during growth and development in the presence of different carbon sources in N. crassa. gpr-4 mutants have reduced mass accumulation compared to the wild type when cultured on high levels of glycerol, mannitol, or arabinose. The defect is most severe on glycerol and is cell density dependent. The genetic and physical relationship between GPR-4 and the three N. crassa G subunits (GNA-1, GNA-2, and GNA-3) was explored. All three G mutants are defective in mass accumulation when cultured on glycerol. However, the phenotypes of gna-1 and gpr-4 gna-1 mutants are identical, introduction of a constitutively activated gna-1 allele suppresses the defects of the gpr-4 mutation, and the carboxy terminus of GPR-4 interacts most strongly with GNA-1 in the yeast two-hybrid assay. Although steady-state cyclic AMP (cAMP) levels are normal in gpr-4 strains, exogenous cAMP partially remediates the dry mass defects of gpr-4 mutants on glycerol medium and gpr-4 strains lack the transient increase in cAMP levels observed in the wild type after addition of glucose to glycerol-grown liquid cultures. Our results support the hypothesis that GPR-4 is coupled to GNA-1 in a cAMP signaling pathway that regulates the response to carbon source in N. crassa. GPR-4-related GPCRs are present in the genomes of several filamentous ascomycete fungal pathogens, raising the possibility that a similar pathway regulates carbon sensing in these organisms.

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* Corresponding author. Mailing address: Department of Plant Pathology, 1415 Boyce Hall, 900 University Avenue, University of California, Riverside, Riverside, CA 92521. Phone: (951) 827-2763. Fax: (951) 827-4294. E-mail: Katherine.Borkovich{at}ucr.edu.

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Eukaryotic Cell, August 2006, p. 1287-1300, Vol. 5, No. 8
1535-9778/06/$08.00+0     doi:10.1128/EC.00109-06
Copyright © 2006, American Society for Microbiology. All Rights Reserved.

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