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Molecular and Cellular Biology, October 2000, p. 7693-7705, Vol. 20, No. 20
0270-7306/00/$04.00+0
Copyright © 2000, American Society for Microbiology. All rights reserved.

Regulation of Conidiation and Adenylyl Cyclase Levels by the Galpha Protein GNA-3 in Neurospora crassa

Ann M. Kays, Patricia S. Rowley, Rudeina A. Baasiri, and Katherine A. Borkovich*

Department of Microbiology and Molecular Genetics, University of Texas--- Houston Medical School, Houston, Texas 77030

Received 17 May 2000/Returned for modification 7 June 2000/Accepted 18 July 2000

We have identified a new gene encoding the G protein alpha  subunit, gna-3, from the filamentous fungus Neurospora crassa. The predicted amino acid sequence of GNA-3 is most similar to the Galpha proteins MOD-D, MAGA, and CPG-2 from the saprophytic fungus Podospora anserina and the pathogenic fungi Magnaporthe grisea and Cryphonectria parasitica, respectively. Deletion of gna-3 leads to shorter aerial hyphae and premature, dense conidiation during growth on solid medium or in standing liquid cultures and to inappropriate conidiation in submerged culture. The conidiation and aerial hypha defects of the Delta gna-3 strain are similar to those of a previously characterized adenylyl cyclase mutant, cr-1. Supplementation with cyclic AMP (cAMP) restores wild-type morphology to Delta gna-3 strains in standing liquid cultures. Solid medium augmented with exogenous cAMP suppresses the premature conidiation defect, but aerial hypha formation is still reduced. Submerged-culture conidiation is refractory to cAMP but is suppressed by peptone. In addition, Delta gna-3 submerged cultures express the glucose-repressible gene, qa-2, to levels greatly exceeding those observed in the wild type under carbon-starved conditions. Delta gna-3 strains exhibit reduced fertility in homozygous crosses during the sexual cycle; exogenous cAMP has no effect on this phenotype. Intracellular steady-state cAMP levels of Delta gna-3 strains are decreased 90% relative to the wild type under a variety of growth conditions. Reduced intracellular cAMP levels in the Delta gna-3 strain correlate with lower adenylyl cyclase activity and protein levels. These results demonstrate that GNA-3 modulates conidiation and adenylyl cyclase levels in N. crassa.

* Corresponding author. Mailing address: Department of Microbiology and Molecular Genetics, University of Texas---Houston Medical School, 6431 Fannin St., Ste. JFB 1.765, Houston, TX 77030. Phone: (713) 500-5438. Fax: (713) 500-5499. E-mail: borkovic{at}utmmg.med.uth.tmc.edu.

Molecular and Cellular Biology, October 2000, p. 7693-7705, Vol. 20, No. 20
0270-7306/00/$04.00+0
Copyright © 2000, American Society for Microbiology. All rights reserved.


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