bio-energetics-investigation-candida-albicans-using-real-time

Bio-energetics Investigation of <em>Candida albicans</em> Using Real-time Extracellular Flux Analysis

Mitochondria are essential organelles for the cellular metabolism and survival. A variety of key events take place in mitochondria, such as cellular ...

The State University of New Jersey

Prokaryotes, eukaryotic microorganisms and plants utilize two-component signal transduction pathways to detect and respond to various environmental cues. These signaling cascades were acquired by eukaryotes via horizontal gene transfer events from ancestral bacteria. Recent exciting discoveries have identified two-component signaling systems in mitochondria and chloroplasts of several eukaryotic microorganisms and plants, therefore providing important clues to the evolutionary transition of these signaling cascades from prokaryotes to eukaryotes. This review will focus on the role of two-component signal transduction pathways in fungal pathogenesis and also discuss key new discoveries of presence of proteins participating in these signaling pathways in mitochondrion. Before addressing these issues, I first briefly describe the magnitude and the economic impact of the healthcare problems caused by fungal pathogens. 

Two-component phosphorelays in fungal mitochondria and beyond.

Morphological plasticity such as the yeast-to-hyphae transition is a key virulence factor of the human fungal pathogen Candida albicans. Hyphal formation is controlled by a multilayer regulatory network composed of environmental sensing, signaling, transcriptional modulators as well as chromatin modifications. Here, we demonstrate a novel role for the replication-independent HIR histone chaperone complex in fungal morphogenesis. HIR operates as a crucial modulator of hyphal development, since genetic ablation of the HIR complex subunit Hir1 decreases sensitivity to morphogenetic stimuli. Strikingly, HIR1-deficient cells display altered transcriptional amplitudes upon hyphal initiation, suggesting that Hir1 affects transcription by establishing transcriptional thresholds required for driving morphogenetic cell-fate decisions. Furthermore, ectopic expression of the transcription factor Ume6, which facilitates hyphal maintenance, rescues filamentation defects of hir1Δ/Δ cells, suggesting that Hir1 impacts the early phase of hyphal initiation. Hence, chromatin chaperone-mediated fine-tuning of transcription is crucial for driving morphogenetic conversions in the fungal pathogen C. albicans.

The Candida albicans HIR histone chaperone regulates the yeast-to-hyphae transition by controlling the sensitivity to morphogenesis signals.

ATP-binding cassette (ABC) transporters help export various substrates across the cell membrane and significantly contribute to drug resistance. However, a recent study reported an unusual case in which the loss of an ABC transporter in , orf19.4531 (previously named ROA1), increases resistance against antifungal azoles, which was attributed to an altered membrane potential in the mutant strain. To obtain further mechanistic insights into this phenomenon, here we confirmed that the plasma membrane-localized transporter (renamed  for consistency with  nomenclature) could efflux xenobiotics such as berberine, rhodamine 123, and paraquat. Moreover, a  null mutant, NKKY101, displayed increased susceptibility to these xenobiotics. Interestingly, fluorescence recovery after photobleaching (FRAP) results indicated that NKKY101 mutant cells exhibited increased plasma membrane rigidity, resulting in reduced azole accumulation and contributing to azole resistance. Transcriptional profiling revealed that ribosome biogenesis genes were significantly up-regulated in the NKKY101 mutant. As ribosome biogenesis is a well-known downstream phenomenon of target of rapamycin (TOR1) signaling, we suspected a link between ribosome biogenesis and TOR1 signaling in NKKY101. Therefore, we grew NKKY101 cells on rapamycin and observed TOR1 hyperactivation, which leads to Hsp90-dependent calcineurin stabilization and thereby increased azole resistance. This  finding was supported by  data from a mouse model of systemic infection in which NKKY101 cells led to higher fungal load after fluconazole challenge than wild-type cells. Taken together, our study uncovers a mechanism of azole resistance in , involving increased membrane rigidity and TOR signaling.

Neeraj Chauhan

Department of Microbiology,
Biochemistry and Molecular Genetics,
New Jersey Medical School,
Rutgers,
Public Health Research Institute,
Department of Microbiology, Biochemistry and Molecular Genetics, New Jersey Medical School, Rutgers,
Public Health Research Institute, Rutgers

The State University of New Jersey

Bio-energetics Investigation of Candida albicans Using Real-time Extracellular Flux Analysis