The Adr1 transcription factor directs regulation of the ergosterol pathway and azole resistance in Candida albicans.

IMPORTANCE: Research often relies on well-studied orthologs within related species, with researchers using a well-studied gene or protein to allow prediction of the function of the ortholog. In the opportunistic pathogen Candida albicans, orthologs are usually compared with Saccharomyces cerevisiae, and this approach has been very fruitful. Many transcription factors (TFs) do similar jobs in the two species, but many do not, and typically changes in function are driven not by modifications in the structures of the TFs themselves but in the connections between the transcription factors and their regulated genes. This strategy of changing TF function has been termed transcription factor rewiring. In this study, we specifically looked for rewired transcription factors, or Candida-specific TFs, that might play a role in drug resistance. We investigated 30 transcription factors that were potentially rewired or were specific to the Candida clade. We found that the Adr1 transcription factor conferred resistance to drugs like fluconazole, amphotericin B, and terbinafine when activated. Adr1 is known for fatty acid and glycerol utilization in Saccharomyces, but our study reveals that it has been rewired and is connected to ergosterol biosynthesis in Candida albicans.

Comparative Evaluation of Compressive and Diametral Tensile Strength in Die Stone Reinforced With Different Types of Nanoparticles: An In Vitro Study.

AIM: To investigate the effect of different types of nanoparticles on the compressive strength (CS) and diametral tensile strength (DTS) of type IV dental stones. MATERIALS AND METHODS: A total of 100 specimens were made from the mould for all five groups. Four commercially available nanoparticles (aluminium oxide (Al2O3), silicon dioxide (SiO2), zinc oxide (ZnO), and zirconium oxide (ZrO2)) were used in this study in a concentration of 10%. CS and DTS tests were performed in a universal test machine. The data were statistically analysed using ANOVA and Student's t-test. RESULTS: The interaction between nanoparticles and the type of dental stone was found to be statistically significant (p < 0.05). CS and DTS values decreased by adding all four nanoparticles. The lowest CS and DTS were observed in 10% ZnO nanoparticles when added to type IV dental stone. CONCLUSION: It was concluded that the addition of nanoparticles (Al2O3, SiO2, ZnO, and ZrO2) to die stone significantly decreased the CS and DTS for all groups. Among all groups, the incorporation of 10% ZrO2 nanoparticles (group E) to die stone showed significantly less decrease in CS and DTS compared to Al2O3, SiO2, and ZnO. Incorporation of ZnO nanoparticles, on the other hand, showed a significantly more amount of decrease in the CS and DTS compared to Al2O3, SiO2, and ZrO2.

Corrigendum to "Loss of Arp1, a putative actin-related protein, triggers filamentous and invasive growth and impairs pathogenicity in Candida albicans" (Computational and Structural Biotechnology Journal, 2020, 18: 4002-4015).

[This corrects the article DOI: 10.1016/j.csbj.2020.11.034.].

Calcium-calcineurin signaling pathway in Candida albicans: A potential drug target.

Increased morbidity and mortality of candidiasis are a notable threat to the immunocompromised patients. At present, the types of drugs available to treat C. albicans infection are relatively limited. Moreover, the emergence of antifungal drug resistance of C. albicans makes the treatment of C. albicans infection more difficult. The calcium-calcineurin signaling pathway plays a crucial role in the survival and pathogenicity of C. albicans and may act as a potential target against C. albicans. In this review, we summarized functions of the calcium-calcineurin signaling pathway in several biological processes, compared the differences of this signaling pathway between C. albicans and humans, and described anti-C. albicans activity of inhibitors of this signaling pathway. We believe that targeting the calcium-calcineurin signaling pathway is a promising strategy to cope with C. albicans infection.

Modulation of the complex regulatory network for methionine biosynthesis in fungi.

The assimilation of inorganic sulfate and the synthesis of the sulfur-containing amino acids methionine and cysteine is mediated by a multibranched biosynthetic pathway. We have investigated this circuitry in the fungal pathogen Candida albicans, which is phylogenetically intermediate between the filamentous fungi and Saccharomyces cerevisiae. In S. cerevisiae, this pathway is regulated by a collection of five transcription factors (Met4, Cbf1, Met28, and Met31/Met32), while in the filamentous fungi the pathway is controlled by a single Met4-like factor. We found that in C. albicans, the Met4 ortholog is also a core regulator of methionine biosynthesis, where it functions together with Cbf1. While C. albicans encodes this Met4 protein, a Met4 paralog designated Met28 (Orf19.7046), and a Met31 protein, deletion, and activation constructs suggest that of these proteins only Met4 is actually involved in the regulation of methionine biosynthesis. Both Met28 and Met31 are linked to other functions; Met28 appears essential, and Met32 appears implicated in the regulation of genes of central metabolism. Therefore, while S. cerevisiae and C. albicans share Cbf1 and Met4 as central elements of the methionine biosynthesis control, the other proteins that make up the circuit in S. cerevisiae are not members of the C. albicans control network, and so the S. cerevisiae circuit likely represents a recently evolved arrangement.

Candida albicans targets that potentially synergize with fluconazole.

Fluconazole has characteristics that make it widely used in the clinical treatment of C. albicans infections. However, fluconazole has only a fungistatic activity in C. albicans, therefore, in the long-term treatment of C. albicans infection with fluconazole, C. albicans has the potential to acquire fluconazole resistance. A promising approach to increase fluconazole's efficacy is identifying potential targets of drugs that can enhance the antifungal effect of fluconazole, or even make the drug fungicidal. In this review, we systematically provide a global overview of potential targets of drugs synergistic with fluconazole in C. albicans, identify new avenues for research on fluconazole potentiation, and highlight the promise of combinatorial strategies with fluconazole in combatting C. albicans infections.

Loss of Arp1, a putative actin-related protein, triggers filamentous and invasive growth and impairs pathogenicity in Candida albicans.

The polymorphous cellular shape of Candida albicans, in particular the transition from a yeast to a filamentous form, is crucial for either commensalism or life-threatening infections of the host. Various external or internal stimuli, including serum and nutrition starvation, have been shown to regulate filamentous growth primarily through two classical signaling pathways, the cAMP-PKA and the MAPK pathways. Genotoxic stress also induces filamentous growth, but through independent pathways, and little is known about negative regulation during this reversible morphological transition. In this study, we established that ARP1 in C. albicans, similar to its homolog in S. cerevisiae, has a role in nuclei separation and spindle orientation. Deletion of ARP1 generated filamentous and invasive growth as well as increased biofilm formation, accompanied by up-regulation of hyphae specific genes, such as HWP1, UME6 and ALS3. The filamentous and invasive growth of the ARP1 deletion strain was independent of transcription factors Efg1, Cph1 and Ume6, but was suppressed by deleting checkpoint BUB2 or overexpressing NRG1. Deletion of ARP1 impaired the colonization of Candida cells in mice and also attenuated virulence in a mouse model. All the data suggest that loss of ARP1 activates filamentous and invasive growth in vitro, and that it positively regulates virulence in vivo, which provides insight into actin-related morphology and pathogenicity in C. albicans.

Hof1 plays a checkpoint-related role in MMS-induced DNA damage response in Candida albicans.

Cells depend on robust DNA damage recognition and repair systems to maintain genomic integrity for survival in a mutagenic environment. In the pathogenic yeast Candida albicans, a subset of genes involved in the response to DNA damage-induced genome instability and morphological changes has been found to regulate virulence. To better understand the virulence-linked DNA repair network, we screened for methyl methane sulfonate (MMS) sensitivity within the GRACE conditional expression collection and identified 56 hits. One of these potential DNA damage repair-associated genes, a HOF1 conditional mutant, unexpectedly had a previously characterized function in cytokinesis. Deletion of HOF1 resulted in MMS sensitivity and genome instability, suggesting Hof1 acts in the DNA damage response. By probing genetic interactions with distinct DNA repair pathways, we found that Hof1 is genetically linked to the Rad53 pathway. Furthermore, Hof1 is down-regulated in a Rad53-dependent manner and its importance in the MMS response is reduced when Rad53 is overexpressed or when RAD4 or RAD23 is deleted. Together, this work expands our understanding of the C. albicans DNA repair network and uncovers interplay between the cytokinesis regulator Hof1 and the Rad53-mediated checkpoint.

Evaluation of Antifungal Efficacy of Three New Cyclic Lipopeptides of the Class Bacillomycin from Bacillus subtilis RLID 12.1.

New lipopeptide homologues (AF3, AF4, and AF5) with antifungal activities against Candida and Cryptococcus spp. were purified from a cell-free supernatant of Bacillus subtilis RLID 12.1. The lipopeptides AF3, AF4, and AF5 were identified with the same peptide sequence Asn-Pro-Tyr-Asn-Gln-Thr-Ser with variations in the fatty acid branching type and chain length (anteiso-C17, iso-C17, and iso-C18, respectively). Upon comparing the three homologues for MICs against 81 Candida (n = 64) and Cryptococcus (n = 17) clinical isolates and their cytotoxicities, we found that AF4 was the most promising antifungal lipopeptide, since it demonstrated 100% inhibition at geometric mean MICs of 3.31, 3.41, 3.48, and 2.83 µg/ml against Candida albicans, Candida tropicalis, Candida auris, and Cryptococcus neoformans, respectively, with low hemolysis values (<6%) and 50% inhibitory concentrations (13.31 µg/ml). The additive effects among the homologues AF3, AF4, and AF5 were evaluated against three Candida species, along with the cytotoxicity studies. Five combinations exhibited good additive interaction effects: AF3/AF4 (at corresponding concentrations of 4 and 4 µg/ml [4/4 µg/ml]), AF3/AF5 (4/4 µg/ml), AF3/AF5 (2/4 µg/ml), AF4/AF5 (4/4 µg/ml), and AF4/AF5 (2/4 µg/ml) in planktonic cell inhibition and AF3/AF4 (4/4 µg/ml), AF3/AF5 (4/4 µg/ml), and AF3/AF5 (2/4 µg/ml) in the inhibition of biofilm formation. However, combinations AF3/AF4 and AF3/AF5, which showed >70% cell survival with low hemolysis (<5%), were found to be comparatively effective. We describe here the additive effects of lipopeptide homologues showing reduced cytotoxicity against mammalian cells; these combinations might serve as a potent antibiofilm-forming substitute.

Enhancement of drought stress tolerance in crops by plant growth promoting rhizobacteria.

Drought is one of the major constraints on agricultural productivity worldwide and is likely to further increase. Several adaptations and mitigation strategies are required to cope with drought stress. Plant growth promoting rhizobacteria (PGPR) could play a significant role in alleviation of drought stress in plants. These beneficial microorganisms colonize the rhizosphere/endo-rhizosphere of plants and impart drought tolerance by producing exopolysaccharides (EPS), phytohormones, 1-aminocyclopropane- 1-carboxylate (ACC) deaminase, volatile compounds, inducing accumulation of osmolytes, antioxidants, upregulation or down regulation of stress responsive genes and alteration in root morphology in acquisition of drought tolerance. The term Induced Systemic Tolerance (IST) was coined for physical and chemical changes induced by microorganisms in plants which results in enhanced tolerance to drought stresses. In the present review we elaborate on the role of PGPR in helping plants to cope with drought stress.