Toward an integrated model of capsule regulation in Cryptococcus neoformans

PLoS Pathog. 2011 Dec;7(12):e1002411. doi: 10.1371/journal.ppat.1002411. Epub 2011 Dec 8.

Abstract

Cryptococcus neoformans is an opportunistic fungal pathogen that causes serious human disease in immunocompromised populations. Its polysaccharide capsule is a key virulence factor which is regulated in response to growth conditions, becoming enlarged in the context of infection. We used microarray analysis of cells stimulated to form capsule over a range of growth conditions to identify a transcriptional signature associated with capsule enlargement. The signature contains 880 genes, is enriched for genes encoding known capsule regulators, and includes many uncharacterized sequences. One uncharacterized sequence encodes a novel regulator of capsule and of fungal virulence. This factor is a homolog of the yeast protein Ada2, a member of the Spt-Ada-Gcn5 Acetyltransferase (SAGA) complex that regulates transcription of stress response genes via histone acetylation. Consistent with this homology, the C. neoformans null mutant exhibits reduced histone H3 lysine 9 acetylation. It is also defective in response to a variety of stress conditions, demonstrating phenotypes that overlap with, but are not identical to, those of other fungi with altered SAGA complexes. The mutant also exhibits significant defects in sexual development and virulence. To establish the role of Ada2 in the broader network of capsule regulation we performed RNA-Seq on strains lacking either Ada2 or one of two other capsule regulators: Cir1 and Nrg1. Analysis of the results suggested that Ada2 functions downstream of both Cir1 and Nrg1 via components of the high osmolarity glycerol (HOG) pathway. To identify direct targets of Ada2, we performed ChIP-Seq analysis of histone acetylation in the Ada2 null mutant. These studies supported the role of Ada2 in the direct regulation of capsule and mating responses and suggested that it may also play a direct role in regulating capsule-independent antiphagocytic virulence factors. These results validate our experimental approach to dissecting capsule regulation and provide multiple targets for future investigation.

Publication types

  • Research Support, N.I.H., Extramural
  • Research Support, Non-U.S. Gov't

MeSH terms

  • Amino Acid Sequence
  • Animals
  • Chromatin Immunoprecipitation
  • Cryptococcosis / genetics
  • Cryptococcosis / metabolism
  • Cryptococcus neoformans / genetics
  • Cryptococcus neoformans / metabolism
  • Cryptococcus neoformans / pathogenicity*
  • Female
  • Fungal Proteins / genetics
  • Fungal Proteins / metabolism*
  • Gene Expression Profiling
  • Gene Expression Regulation, Fungal
  • Histone Acetyltransferases / genetics
  • Histone Acetyltransferases / metabolism*
  • Mice
  • Mice, Inbred C57BL
  • Microscopy, Fluorescence
  • Molecular Sequence Data
  • Polysaccharides / genetics
  • Polysaccharides / metabolism*
  • Sequence Homology, Amino Acid
  • Transcription, Genetic
  • Virulence / genetics
  • Virulence Factors / genetics
  • Virulence Factors / metabolism

Substances

  • Fungal Proteins
  • Polysaccharides
  • Virulence Factors
  • Histone Acetyltransferases