This proposal will determine the molecular basis for adhesion of fungi, Saccharomyces and Candida species to inert surfaces. Our goal is to solve both basic and clinically relevant problems as fungal pathogens gain entry into the body by adhesion to indwelling devices. Prevention of adhesion to these devices could stem the increasing prevalence of fungal disease. Adhesion in fungi is conferred by a family of genes that encode cell surface proteins. The basic studies take advantage of our ability to compare the functions of all the genes in the genomes of two closely related strains of S. cerevisiae (S288c and sigma) with a specific emphasis on those genes that affect adhesion. This comparison is possible because the genome of sigma will be sequenced and annotated in concert with the transfer of all the deletion set (~6000 knockouts) from the already sequenced S288c into sigma. Reciprocally, the library of S288c deletions has been transformed with FLO8+ to permit assessment of adhesion in this background. This comparison will reveal all genes required for the adhesion phenotype, and also provide evidence for the regulatory divergence between individuals within an interbreeding population. Variegation, the on/off switching of telomeric and non-telomeric adhesin genes, will be studied by continuous fluorescence microscopy. The cis-acting sequences and trans-acting complexes that control expression and variegation will be analyzed. This analysis will reveal the components of required for establishment and maintenance of this epigenetic state. The control of gene expression by antisense RNA and small open reading frames in the promoters of genes will be determined using bioinformatics as well as molecular genetics. This information obtained from Saccharomyces is used to create strains of C. albicans for use in devising dextran gels containing non-diffusible amphotericin. The Amphogels are effective in an animal model. New work will devise gels that are effective against adherent, non-adherent, and filamentous forms of C. albicans and other fungal species.

Public Health Relevance

Fungi, a major source of human disease, gain entry into the body mainly by adhering to medical devices such as catheters. The research proposed will determine the mechanism of adherence by fungi. This basic information will guide the creation of antibiotic containing materials that can be used to coat devices so that the fungi are killed upon contact before they can hitch a ride into the body.

Agency
National Institute of Health (NIH)
Institute
National Institute of General Medical Sciences (NIGMS)
Type
Research Project (R01)
Project #
5R01GM035010-28
Application #
8197675
Study Section
Molecular Genetics B Study Section (MGB)
Program Officer
Reddy, Michael K
Project Start
1984-07-01
Project End
2013-09-14
Budget Start
2011-12-01
Budget End
2013-09-14
Support Year
28
Fiscal Year
2012
Total Cost
$738,758
Indirect Cost
$359,908
Name
Whitehead Institute for Biomedical Research
Department
Type
DUNS #
120989983
City
Cambridge
State
MA
Country
United States
Zip Code
02142
Vyas, Valmik K; Barrasa, M Inmaculada; Fink, Gerald R (2015) A Candida albicans CRISPR system permits genetic engineering of essential genes and gene families. Sci Adv 1:e1500248
Lam, Felix H; Ghaderi, Adel; Fink, Gerald R et al. (2014) Biofuels. Engineering alcohol tolerance in yeast. Science 346:71-5
Edwards, Matthew D; Symbor-Nagrabska, Anna; Dollard, Lindsey et al. (2014) Interactions between chromosomal and nonchromosomal elements reveal missing heritability. Proc Natl Acad Sci U S A 111:7719-22
Wang, Benjamin L; Ghaderi, Adel; Zhou, Hang et al. (2014) Microfluidic high-throughput culturing of single cells for selection based on extracellular metabolite production or consumption. Nat Biotechnol 32:473-8
Schwartz, Schraga; Bernstein, Douglas A; Mumbach, Maxwell R et al. (2014) Transcriptome-wide mapping reveals widespread dynamic-regulated pseudouridylation of ncRNA and mRNA. Cell 159:148-62
Schwartz, Schraga; Agarwala, Sudeep D; Mumbach, Maxwell R et al. (2013) High-resolution mapping reveals a conserved, widespread, dynamic mRNA methylation program in yeast meiosis. Cell 155:1409-21
Avalos, José L; Fink, Gerald R; Stephanopoulos, Gregory (2013) Compartmentalization of metabolic pathways in yeast mitochondria improves the production of branched-chain alcohols. Nat Biotechnol 31:335-41
Bumgarner, Stacie L; Neuert, Gregor; Voight, Benjamin F et al. (2012) Single-cell analysis reveals that noncoding RNAs contribute to clonal heterogeneity by modulating transcription factor recruitment. Mol Cell 45:470-82
Agarwala, Sudeep D; Blitzblau, Hannah G; Hochwagen, Andreas et al. (2012) RNA methylation by the MIS complex regulates a cell fate decision in yeast. PLoS Genet 8:e1002732
Ryan, Owen; Shapiro, Rebecca S; Kurat, Christoph F et al. (2012) Global gene deletion analysis exploring yeast filamentous growth. Science 337:1353-6

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