Structure-Guided Discovery of Potent Antifungals that Prevent Ras Signaling by Inhibiting Protein Farnesyltransferase

Journal of Medicinal Chemistry · 2022 · 12 citations

• Chemistry
• Biochemistry
• Pharmacology

levels of enzyme and farnesyl substrate. We elucidated how chemical modifications of the antifungals encode desired inhibitor conformation and concomitant inhibitory mechanism.

Comparative analysis of RNA enrichment methods for preparation of <i>Cryptococcus neoformans</i> RNA sequencing libraries

G3 Genes Genomes Genetics · 2021 · 12 citations

• Biology
• Molecular biology
• Computational biology

RNA sequencing (RNA-Seq) experiments focused on gene expression involve removal of ribosomal RNA (rRNA) because it is the major RNA constituent of cells. This process, called RNA enrichment, is done primarily to reduce cost: without rRNA removal, deeper sequencing must be performed to compensate for the sequencing reads wasted on rRNA. The ideal RNA enrichment method removes all rRNA without affecting other RNA in the sample. We tested the performance of three RNA enrichment methods on RNA isolated from Cryptococcus neoformans, a fungal pathogen of humans. We find that the RNase H depletion method is more efficient in depleting rRNA and more specific in recapitulating non-rRNA levels present in unenriched controls than the commonly-used Poly(A) isolation method. The RNase H depletion method is also more effective than the Ribo-Zero depletion method as measured by rRNA depletion efficiency and recapitulation of protein-coding RNA levels present in unenriched controls, while the Ribo-Zero depletion method more closely recapitulates annotated non-coding RNA (ncRNA) levels. Finally, we leverage these data to accurately map the C. neoformans mitochondrial rRNA genes, and also demonstrate that RNA-Seq data generated with the RNase H and Ribo-Zero depletion methods can be used to explore novel C. neoformans long non-coding RNA genes.

Transposon mobilization in the human fungal pathogen <i>Cryptococcus</i> is mutagenic during infection and promotes drug resistance in vitro

Proceedings of the National Academy of Sciences · 2020 · 71 citations

• Biology
• Microbiology
• Genetics

is an important mechanism that enhances microbial adaptation and promotes pathogenesis and drug resistance in the human host.

Erg6 affects membrane composition and virulence of the human fungal pathogen Cryptococcus neoformans

Fungal Genetics and Biology · 2020 · 56 citations

• Biology
• Microbiology
• Biochemistry

Length Specificity and Polymerization Mechanism of (1,3)-β-<scp>d</scp>-Glucan Synthase in Fungal Cell Wall Biosynthesis

Biochemistry · 2020 · 32 citations

• Biochemistry
• Chemistry
• Stereochemistry

, which represents the first observation of chain elongation by a nucleotide-sugar-dependent polysaccharide synthase. Coupling the SEC-PAD-RC method with substrate analogue mechanistic probes provided the first unambiguous evidence that GS catalyzes non-reducing end polymerization. On the basis of these observations, we propose a detailed model for the catalytic mechanism of GS. The approaches described here can be used to determine the mechanism of catalysis of other polysaccharide synthases.