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Molecular and General Genetics MGG

, Volume 256, Issue 4, pp 426–435 | Cite as

Fil1, a G-protein α-subunit that acts upstream of cAMP and is essential for dimorphic switching in haploid cells of Ustilago hordei

  • A. Lichter
  • D. Mills
ORIGINAL PAPER
  • 35 Downloads

Abstract

A constitutive mutation, fil1, that causes filamentous growth in the haplophase of the dimorphic smut fungus Ustilago hordei, was previously shown to be genetically associated with a 50-kb deletion within a 940-kb chromosome. Physiological studies suggested that a gene that functions upstream of adenylyl cyclase was deleted in the mutant. Representational difference analysis of isolated chromosomes was used to obtain deletion-specific DNA probes and corresponding genomic cosmid clones. Complementation analysis identified a cosmid clone and subsequently a 2.1-kb insert that converted transformants of the mutant strain10.1a(fil1) from the filamentous to the sporidial cell type. A single open reading frame of 354 codons that encodes a putative α-subunit of the heterotrimeric G-proteins was identified. Fil1 displayed a high degree of sequence identity to Gpa1 from the basidiomycete Cryptococcus neoformans and CPG-2 from the ascomycete Cryphonectria parasitica. FIL1, when introduced on a self-replicating vector, was found to suppress filamentous growth of starved haploid wild-type strains and restore normal mating response to the fil1 mutant, but did not suppress sexual dimorphism of either strain. Fil1 appears to function analogously to mammalian Gα proteins, which are coupled to cAMP production via adenylyl cyclase, to regulate dimorphic switching in  U. hordei.

Key words Chromosomal deletion Heterotrimeric G-proteins Fungal dimorphism cAMP Representational difference analysis (RDA) 

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Copyright information

© Springer-Verlag Berlin Heidelberg 1997

Authors and Affiliations

  • A. Lichter
    • 1
  • D. Mills
    • 1
  1. 1.Department of Botany and Plant Pathology, Oregon State University, 2082 Cordley Hall, Corvallis, OR 97331-2902, USA Fax: +1-541-737-3573; e-mail: millsd@bcc.orst.eduUS

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