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Eukaryotic Cell, September 2006, p. 1503-1516, Vol. 5, No. 9
1535-9778/06/$08.00+0     doi:10.1128/EC.00124-06
Copyright © 2006, American Society for Microbiology. All Rights Reserved.

The Predicted G-Protein-Coupled Receptor GPR-1 Is Required for Female Sexual Development in the Multicellular Fungus Neurospora crassa

Svetlana Krystofova and Katherine A. Borkovich*

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

Received 28 April 2006/ Accepted 30 June 2006

G-protein-coupled receptors (GPCRs) control important aspects of asexual and sexual development in eukaryotic organisms. We have identified a predicted GPCR in the filamentous fungus Neurospora crassa with similarity to cyclic AMP-receptor like GPCRs from Dictyostelium discoideum and GCR1 from Arabidopsis thaliana. Expression of gpr-1 is highest in female reproductive structures, and deletion of gpr-1 leads to defects during sexual development. Unfertilized female structures (protoperithecia) from {Delta}gpr-1 strains are weakly pigmented, small, and submerged in the agar. The perithecia produced after fertilization have deformed beaks that lack ostioles, the openings through which ascospores are discharged. Localization studies using a GPR-1-green fluorescent protein fusion protein showed that GPR-1 is targeted to female reproductive structures. Genetic epistasis experiments with the three G{alpha} genes were inconclusive due to the early block in mating exhibited by {Delta}gna-1 strains. Phenotypic analysis of mutants from a high-throughput N. crassa knockout project allowed identification of BEK-1, a homeodomain transcription factor that is a potential target of GPR-1. The perithecial defects of {Delta}bek-1 strains are similar to those of the {Delta}gpr-1 strain, and epistasis analysis indicates that bek-1 could function downstream of gpr-1 during postfertilization events. The effect must be posttranscriptional, as bek-1 transcript levels are not affected in {Delta}gpr-1 strains. The lack of ostioles in {Delta}gpr-1 and {Delta}bek-1 mutants has an undesirable effect on the ability to spread progeny (ascospores) by the normal ejection mechanism and would severely compromise the fitness of these strains in nature.

* Corresponding author. Mailing address: Department of Plant Pathology, University of California, 1415 Boyce Hall, 900 University Ave., Riverside, CA 92521. Phone: (951) 827-2753. Fax: (951) 827-4294. E-mail: Katherine.Borkovich{at}ucr.edu.

Eukaryotic Cell, September 2006, p. 1503-1516, Vol. 5, No. 9
1535-9778/06/$08.00+0     doi:10.1128/EC.00124-06
Copyright © 2006, American Society for Microbiology. All Rights Reserved.



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