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Eukaryotic Cell, January 2006, p. 103-111, Vol. 5, No. 1
1535-9778/06/$08.00+0     doi:10.1128/EC.5.1.103-111.2006
Copyright © 2006, American Society for Microbiology. All Rights Reserved.

Cryptococcus neoformans Senses CO2 through the Carbonic Anhydrase Can2 and the Adenylyl Cyclase Cac1

Estelle Gewiss Mogensen,1,{dagger} Guilhem Janbon,2,{dagger} James Chaloupka,3 Clemens Steegborn,4 Man Shun Fu,1 Frédérique Moyrand,2 Torsten Klengel,1 David S. Pearson,1 Michael A. Geeves,1 Jochen Buck,3 Lonny R. Levin,3 and Fritz A. Mühlschlegel1*

Department of Biosciences, University of Kent, Canterbury, Kent CT2 7NJ, United Kingdom,1 Unité de Mycologie Moléculaire, CNRS FRE2849, Institut Pasteur, 25 rue du Dr. Roux, 75015 Paris, France,2 Department of Pharmacology, Joan and Sanford I. Weill Medical College of Cornell University, New York, New York 10021,3 Department of Physiological Chemistry, Ruhr-Universität Bochum, Universitätsstrasse 150, 44801 Bochum, Germany4

Received 17 October 2005/ Accepted 18 October 2005

Cryptococcus neoformans, a fungal pathogen of humans, causes fatal meningitis in immunocompromised patients. Its virulence is mainly determined by the elaboration of a polysaccharide capsule surrounding its cell wall. During its life, C. neoformans is confronted with and responds to dramatic variations in CO2 concentrations; one important morphological change triggered by the shift from its natural habitat (0.033% CO2) to infected hosts (5% CO2) is the induction of capsule biosynthesis. In cells, CO2 is hydrated to bicarbonate in a spontaneous reaction that is accelerated by carbonic anhydrases. Here we show that C. neoformans contains two ß-class carbonic anhydrases, Can1 and Can2. We further demonstrate that CAN2, but not CAN1, is abundantly expressed and essential for the growth of C. neoformans in its natural environment, where CO2 concentrations are limiting. Structural studies reveal that Can2 forms a homodimer in solution. Our data reveal Can2 to be the main carbonic anhydrase and suggest a physiological role for bicarbonate during C. neoformans growth. Bicarbonate directly activates the C. neoformans Cac1 adenylyl cyclase required for capsule synthesis. We show that this specific activation is optimal at physiological pH.

* Corresponding author. Mailing address: Department of Biosciences, University of Kent, Canterbury, Kent CT2 7NJ, United Kingdom. Phone: 44 1227 823988. Fax: 44 1227 763912. E-mail: F.A.Muhlschlegel{at}kent.ac.uk.

{dagger} These authors contributed equally to this work.

Eukaryotic Cell, January 2006, p. 103-111, Vol. 5, No. 1
1535-9778/06/$08.00+0     doi:10.1128/EC.5.1.103-111.2006
Copyright © 2006, American Society for Microbiology. All Rights Reserved.



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