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Eukaryotic Cell, February 2005, p. 407-420, Vol. 4, No. 2
1535-9778/05/$08.00+0     doi:10.1128/EC.4.2.407-420.2005
Copyright © 2005, American Society for Microbiology. All Rights Reserved.

The Function of the Coding Sequences for the Putative Pheromone Precursors in Podospora anserina Is Restricted to Fertilization

Evelyne Coppin,* Christelle de Renty, and Robert Debuchy

Institut de Génétique et Microbiologie, UMR 8621 CNRS, Université Paris Sud, Orsay, France

Received 24 August 2004/ Accepted 9 December 2004

We cloned the pheromone precursor genes of Podospora anserina in order to elucidate their role in the biology of this fungus. The mfp gene encodes a 24-amino-acid polypeptide finished by the CAAX motif, characteristic of fungal lipopeptide pheromone precursors similar to the a-factor precursor of Saccharomyces cerevisiae. The mfm gene encodes a 221-amino-acid polypeptide, which is related to the S. cerevisiae {alpha}-factor precursor and contains two 13-residue repeats assumed to correspond to the mature pheromone. We deleted the mfp and mfm coding sequence by gene replacement. The mutations specifically affect male fertility, without impairing female fertility and vegetative growth. The male defect is mating type specific: the mat+ {Delta}mfp and mat– {Delta}mfm mutants produce male cells inactive in fertilization whereas the mat– {Delta}mfp and mat+ {Delta}mfm mutants show normal male fertility. Genetic data indicate that both mfp and mfm are transcribed at a low level in mat+ and mat– vegetative hyphae. Northern-blot analysis shows that their transcription is induced by the mating types in microconidia (mfp by mat+ and mfm by mat–). We managed to cross {Delta}mfp {Delta}mfm strains of opposite mating type, by complementation and transient expression of the pheromone precursor gene to trigger fertilization. These crosses were fertile, demonstrating that once fertilization occurs, the pheromone precursor genes are unnecessary for the completion of the sexual cycle. Finally, we show that the constitutively transcribed gpd::mfm and gpd::mfp constructs are repressed at a posttranscriptional level by the noncognate mating type.

* Corresponding author. Mailing address: IGM Batiment 400, Université Paris-Sud, F-91405 Orsay Cedex, France. Phone: 33 1 69 15 70 12. Fax: 33 1 69 15 70 06. E-mail: evelyne.coppin{at}igmors.u-psud.fr.

Eukaryotic Cell, February 2005, p. 407-420, Vol. 4, No. 2
1535-9778/05/$08.00+0     doi:10.1128/EC.4.2.407-420.2005
Copyright © 2005, American Society for Microbiology. All Rights Reserved.



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