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Eukaryotic Cell, May 2007, p. 831-843, Vol. 6, No. 5
1535-9778/07/$08.00+0     doi:10.1128/EC.00269-06
Copyright © 2007, American Society for Microbiology. All Rights Reserved.

The WW Domain Protein PRO40 Is Required for Fungal Fertility and Associates with Woronin Bodies{triangledown} ,{dagger}

Ines Engh,1 Christian Würtz,2 Konstanze Witzel-Schlömp,1 Hai Yu Zhang,1 Birgit Hoff,1 Minou Nowrousian,1 Hanspeter Rottensteiner,2 and Ulrich Kück1*

Lehrstuhl für Allgemeine und Molekulare Botanik,1 Institut für Physiologische Chemie, Abteilung für Systembiochemie, Ruhr-Universität Bochum, D-44780 Bochum, Germany2

Received 22 August 2006/ Accepted 27 February 2007

Fruiting body formation in ascomycetes is a highly complex process that is under polygenic control and is a fundamental part of the fungal sexual life cycle. However, the molecular determinants regulating this cellular process are largely unknown. Here we show that the sterile pro40 mutant is defective in a 120-kDa WW domain protein that plays a pivotal role in fruiting body maturation of the homothallic ascomycete Sordaria macrospora. Although WW domains occur in many eukaryotic proteins, homologs of PRO40 are present only in filamentous ascomycetes. Complementation analysis with different pro40 mutant strains, using full-sized or truncated versions of the wild-type pro40 gene, revealed that the C terminus of PRO40 is crucial for restoring the fertile phenotype. Using differential centrifugation and protease protection assays, we determined that a PRO40-FLAG fusion protein is located within organelles. Further microscopic investigations of fusion proteins with DsRed or green fluorescent protein polypeptides showed a colocalization of PRO40 with HEX-1, a Woronin body-specific protein. However, the integrity of Woronin bodies is not affected in mutant strains of S. macrospora and Neurospora crassa, as shown by fluorescence microscopy, sedimentation, and immunoblot analyses. We discuss the function of PRO40 in fruiting body formation.

* Corresponding author. Mailing address: Lehrstuhl für Allgemeine und Molekulare Botanik, Ruhr-Universität Bochum, ND7/131, Universitätsstrasse 150, 44780 Bochum, Germany. Phone: 49-234-3226212. Fax: 49-234-3214184. E-mail: ulrich.kueck{at}rub.de

{triangledown} Published ahead of print on 9 March 2007.

{dagger} Supplemental material for this article may be found at http://ec.asm.org/.

Eukaryotic Cell, May 2007, p. 831-843, Vol. 6, No. 5
1535-9778/07/$08.00+0     doi:10.1128/EC.00269-06
Copyright © 2007, American Society for Microbiology. All Rights Reserved.



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