UDP-glucose dehydrogenase plays multiple roles in the biology of the pathogenic fungus Cryptococcus neoformans

Cara L Griffith; J Stacey Klutts; Lijuan Zhang; Steven B Levery; Tamara L Doering

doi:10.1074/jbc.M408889200

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UDP-glucose dehydrogenase plays multiple roles in the biology of the pathogenic fungus Cryptococcus neoformans

Journal article

Open access

Peer reviewed

UDP-glucose dehydrogenase plays multiple roles in the biology of the pathogenic fungus Cryptococcus neoformans

Cara L Griffith, J Stacey Klutts, Lijuan Zhang, Steven B Levery and Tamara L Doering

The Journal of biological chemistry, Vol.279(49), pp.51669-51676

12/03/2004

DOI: 10.1074/jbc.M408889200

PMID: 15383535

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Abstract

Cryptococcus neoformans is a pathogenic fungus surrounded by an elaborate polysaccharide capsule that is strictly required for its virulence in humans and other mammals. Nearly half of the sugar residues in the capsule are derived from UDP-glucuronic acid or its metabolites. To examine the role of these nucleotide sugars in C. neoformans, the gene encoding UDP-glucose dehydrogenase was disrupted. Mass spectrometry analysis of nucleotide sugar pools showed that the resulting mutant lacked both UDP-glucuronic acid and its downstream product, UDP-xylose, thus confirming the effect of the knockout and indicating that an alternate pathway for UDP-glucuronic acid production was not used. The mutant was dramatically affected by the lack of specific sugar donors, demonstrating altered cell integrity, temperature sensitivity, lack of growth in an animal model of cryptococcosis, and morphological defects. Additionally, the polysaccharide capsule could not be detected on the mutant cells, although the possibility remains that abbreviated forms of capsule components are made, possibly without proper surface display. The capsule defect is largely independent of the other observed changes, as cells that are acapsular because of mutations in other genes show lack of virulence but do not exhibit alterations in cell integrity, temperature sensitivity, or cellular morphology. All of the observed alterations were reversed by correction of the gene disruption.

Microscopy, Electron, Transmission

Cryptococcus neoformans - physiology

Temperature

Coloring Agents - pharmacology

Immunoblotting

Chromatography, High Pressure Liquid

Microscopy, Electron

Microscopy, Interference

Genetic Complementation Test

Uridine Diphosphate Glucose Dehydrogenase - physiology

Nucleotides - chemistry

DNA - chemistry

Uridine Diphosphate Glucose Dehydrogenase - genetics

Mass Spectrometry

Uridine Diphosphate Xylose - chemistry

Polysaccharides - chemistry

Protein Binding

Cell Membrane - metabolism

Models, Genetic

Mutation

Glucuronic Acid - chemistry

Antibodies, Monoclonal - chemistry

Microscopy, Fluorescence

Details

Title: Subtitle: UDP-glucose dehydrogenase plays multiple roles in the biology of the pathogenic fungus Cryptococcus neoformans
Creators: Cara L Griffith - Department of Molecular Microbiology, Washington University School of Medicine, St. Louis, Missouri 63110, USA
J Stacey Klutts
Lijuan Zhang
Steven B Levery
Tamara L Doering
Resource Type: Journal article
Publication Details: The Journal of biological chemistry, Vol.279(49), pp.51669-51676
DOI: 10.1074/jbc.M408889200
PMID: 15383535
NLM abbreviation: J Biol Chem
ISSN: 0021-9258
eISSN: 1083-351X
Publisher: United States
Grant note: R01 GM066303 / NIGMS NIH HHS GM R01 069968 / NIGMS NIH HHS GM R01 066303 / NIGMS NIH HHS P20 RR16459 / NCRR NIH HHS GM F32 072341 / NIGMS NIH HHS
Language: English
Date published: 12/03/2004
Academic Unit: Pathology
Record Identifier: 9984046831102771

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