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Eukaryotic Cell, February 2005, p. 365-378, Vol. 4, No. 2
1535-9778/05/$08.00+0     doi:10.1128/EC.4.2.365-378.2005
Copyright © 2005, American Society for Microbiology. All Rights Reserved.

The Heterotrimeric G-Protein Subunits GNG-1 and GNB-1 Form a Gß{gamma} Dimer Required for Normal Female Fertility, Asexual Development, and G{alpha} Protein Levels in Neurospora crassa

Svetlana Krystofova and Katherine A. Borkovich*

Department of Plant Pathology, University of California—Riverside, Riverside, California

Received 24 September 2004/ Accepted 18 November 2004

We have identified a gene encoding a heterotrimeric G protein {gamma} subunit, gng-1, from the filamentous fungus Neurospora crassa. gng-1 possesses a gene structure similar to that of mammalian G{gamma} genes, consisting of three exons and two introns, with introns present in both the open reading frame and 5'-untranslated region. The GNG-1 amino acid sequence displays high identity to predicted G{gamma} subunits from other filamentous fungi, including Giberella zeae, Cryphonectria parasitica, Trichoderma harzianum, and Magnaporthe grisea. Deletion of gng-1 leads to developmental defects similar to those previously characterized for {Delta}gnb-1 (Gß) mutants. {Delta}gng-1, {Delta}gnb-1, and {Delta}gng-1 {Delta}gnb-1 strains conidiate inappropriately in submerged cultures and are female sterile, producing aberrant female reproductive structures. Similar to previous results obtained with {Delta}gnb-1 mutants, loss of gng-1 negatively influences levels of G{alpha} proteins (GNA-1, GNA-2, and GNA-3) in plasma membrane fractions isolated from various tissues of N. crassa and leads to a significant reduction in the amount of intracellular cyclic AMP. In addition, we show that GNB-1 is essential for maintenance of normal steady-state levels of GNG-1, suggesting a functional interaction between GNB-1 and GNG-1. Direct evidence for a physical association between GNB-1 and GNG-1 in vivo was provided by coimmunoprecipitation.

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

Eukaryotic Cell, February 2005, p. 365-378, Vol. 4, No. 2
1535-9778/05/$08.00+0     doi:10.1128/EC.4.2.365-378.2005
Copyright © 2005, American Society for Microbiology. All Rights Reserved.



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