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Eukaryotic Cell, September 2006, p. 1490-1502, Vol. 5, No. 9
1535-9778/06/$08.00+0     doi:10.1128/EC.00113-06
Copyright © 2006, American Society for Microbiology. All Rights Reserved.

A Eukaryote without Catalase-Containing Microbodies: Neurospora crassa Exhibits a Unique Cellular Distribution of Its Four Catalases{dagger}

Wolfgang Schliebs,1,{ddagger} Christian Würtz,1,{ddagger} Wolf-Hubert Kunau,1 Marten Veenhuis,2 and Hanspeter Rottensteiner1*

Institut für Physiologische Chemie, Abt. Systembiochemie, Ruhr-Universität Bochum, 44780 Bochum, Germany,1 Institute of Biology, University of Groningen, 9751 NN Haren, The Netherlands2

Received 19 April 2006/ Accepted 5 July 2006

Microbodies usually house catalase to decompose hydrogen peroxide generated within the organelle by the action of various oxidases. Here we have analyzed whether peroxisomes (i.e., catalase-containing microbodies) exist in Neurospora crassa. Three distinct catalase isoforms were identified by native catalase activity gels under various peroxisome-inducing conditions. Subcellular fractionation by density gradient centrifugation revealed that most of the spectrophotometrically measured activity was present in the light upper fractions, with an additional small peak coinciding with the peak fractions of HEX-1, the marker protein for Woronin bodies, a compartment related to the microbody family. However, neither in-gel assays nor monospecific antibodies generated against the three purified catalases detected the enzymes in any dense organellar fraction. Furthermore, staining of an N. crassa wild-type strain with 3,3'-diaminobenzidine and H2O2 did not lead to catalase-dependent reaction products within microbodies. Nonetheless, N. crassa does possess a gene (cat-4) whose product is most similar to the peroxisomal type of monofunctional catalases. This novel protein indeed exhibited catalase activity, but was not localized to microbodies either. We conclude that N. crassa lacks catalase-containing peroxisomes, a characteristic that is probably restricted to a few filamentous fungi that produce little hydrogen peroxide within microbodies.

* Corresponding author. Mailing address: Institut für Physiologische Chemie, Abt. Systembiochemie, Ruhr-Universität Bochum, D-44780 Bochum, Germany. Phone: 49-234-322-7046. Fax: 49-234-321-4266. E-mail: Hanspeter.Rottensteiner{at}rub.de.

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

{ddagger} W.S. and C.W. contributed equally to this study.

Eukaryotic Cell, September 2006, p. 1490-1502, Vol. 5, No. 9
1535-9778/06/$08.00+0     doi:10.1128/EC.00113-06
Copyright © 2006, American Society for Microbiology. All Rights Reserved.



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