Transcription factor CCG-8 plays a pivotal role in azole adaptive responses of Neurospora crassa by regulating intracellular azole accumulation

Xue, Wei; Yin, Yunqiang; Ismail, Fatima; Hu, Chengcheng; Zhou, Mi; Cao, Xianhe; Li, Shaojie; Sun, Xiepeng

doi:10.1007/s00294-018-0924-7

Cited by 9 publications

(6 citation statements)

References 43 publications

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“…This created a statistical excess ( p = 10 −4 for motifs with scores ≤2.0) of far less statistical significance than for Osh3 ( p = 10 −26 ) or Osh1 ( p = 10 −44 ). Many of the Opi1 orthologues have an alternate name: “related to clock-controlled protein 8 (ccg-8).” ccg-8 is a Neurospora crassa transcription factor that regulates an aspect of lipid metabolism different from that regulated by Opi1p in S. cerevisiae (Xue et al., 2019). FFAT motifs are absent from all proteins named ccg-8.…”

Section: Resultsmentioning

confidence: 99%

Systematic Prediction of FFAT Motifs Across Eukaryote Proteomes Identifies Nucleolar and Eisosome Proteins With the Predicted Capacity to Form Bridges to the Endoplasmic Reticulum

Slee¹,

Levine²

2019

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The endoplasmic reticulum (ER), the most pervasive organelle, exchanges information and material with many other organelles, but the extent of its inter-organelle connections and the proteins that form bridges are not well known. The integral ER membrane protein VAMP-associated protein (VAP) is found in multiple bridges, interacting with many proteins that contain a short linear motif consisting of “two phenylalanines in an acidic tract” (FFAT). The VAP-FFAT interaction is the most common mechanism by which cytoplasmic proteins, particularly inter-organelle bridges, target the ER. Therefore, predicting new FFAT motifs may both find new individual peripheral ER proteins and identify new routes of communication involving the ER. Here we searched for FFAT motifs across whole proteomes. The excess of eukaryotic proteins with FFAT motifs over background was ≥0.8%, suggesting this is the minimum number of peripheral ER proteins. In yeast, where VAP was previously known to bind 4 proteins with FFAT motifs, a detailed analysis of a subset of proteins predicted 20 FFAT motifs. Extrapolating these findings to the whole proteome estimated the number of FFAT motifs in yeast at approximately 50-55 (0.9% of proteome). Among these previously unstudied FFAT motifs, most have known functions outside the ER, so could be involved in inter-organelle communication. Many of these can target well-characterised membrane contact sites, however some are in nucleoli and eisosomes, organelles previously unknown to have molecular bridges to the ER. We speculate that the nucleolar and eisosomal proteins with predicted motifs may function while bridging to the ER, indicating novel ER-nucleolus and ER-eisosome routes of inter-organelle communication.

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Section: Resultsmentioning

confidence: 99%

Systematic Prediction of FFAT Motifs Across Eukaryote Proteomes Identifies Nucleolar and Eisosome Proteins With the Predicted Capacity to Form Bridges to the Endoplasmic Reticulum

Slee¹,

Levine²

2019

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“…Because deletion of ccg-8 in N. crassa also resulted in azole accumulation and attenuated responses of erg genes to azoles similar to what was seen with the stk-17 KO strain ( 6 , 31 ), we examined the relationship between STK-17 and the transcription factor CCG-8. However, the double-deletion mutant of stk-17 and ccg-8 was more sensitive than either of the single-deletion mutants (Fig.…”

Section: Resultsmentioning

confidence: 99%

“…Azole accumulation is largely affected by efflux pump activity ( 2 ), and CDR4 is the primary azole efflux pump in N. crassa ( 30 – 32 ). To determine if CDR4-mediated azole efflux affected the accumulation of KTC in the stk-17 KO strain, we compared the expression of cdr4 in WT and the stk-17 KO strain.…”

Section: Resultsmentioning

confidence: 99%

Coordinated Regulation of Membrane Homeostasis and Drug Accumulation by Novel Kinase STK-17 in Response to Antifungal Azole Treatment

Zhou

Cao

et al. 2022

Microbiol Spectr

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“…The wild-type (WT) strain FGSC#4200 and the deletion mutant of cdr4 (cdr4 KO , FGSC#11238) were obtained from Fungal Genetic Stock Center (FGSC). The overexpression strain of cdr4 was constructed using the plasmid pCB1532-Pcfp-cdr4-TrpC [32]. Briefly, the plasmid was transformed into the WT strain by electroporation [34].…”

Section: Strains and Cultural Conditionsmentioning

confidence: 99%

“…The novel transcription factor ADS-1, positively regulating the transcriptional responses of azole efflux pumps and targeting genes both in Aspergillus flavus and Fusarium verlicillioides, was found and characterized for the first time in N. crassa [29]. Moreover, the functions of ads-4, csp-1 ccg8, and stk-17, which regulate the adaptive responses to antifungal azoles, were found in N. crassa and were demonstrated to be functionally conserved in pathogenic fungi [30][31][32][33]. Thus, N. crassa has advantages in the study mechanisms of antifungal resistance, particularly in filamentous molds.…”

Section: Introductionmentioning

confidence: 96%

Experimental Evolution of Multidrug Resistance in Neurospora crassa under Antifungal Azole Stress

Zhou

Yin

et al. 2022

JoF

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Multidrug resistance, defined as the resistance to multiple drugs in different categories, has been an increasing serious problem. Limited antifungal drugs and the rapid emergence of antifungal resistance prompt a thorough understanding of how the occurrence of multidrug resistance develops and which mechanisms are involved. In this study, experimental evolution was performed under single-azole-drug stress with the model filamentous fungus Neurospora crassa. By about 30 weeks of continuous growth on agar plates containing ketoconazole or voriconazole with weekly transfer, four evolved multidrug-resistant strains 30thK1, 30thK2, 26thV1, and 24thV2 were obtained. Compared to the ancestral strain, all four strains increased resistance not only to commonly used azoles, including ketoconazole, voriconazole, itraconazole, fluconazole, and triadimefon, but also to antifungal drugs in other categories, including terbinafine (allylamine), amorolfine (morpholine), amphotericin B (polyene), polyoxin B (chitin synthesis inhibitor), and carbendazim (β-tubulin inhibitor). After 8 weeks of growth on agar plates without antifungal drugs with weekly transfer, these evolved strains still displayed multidrug-resistant phenotype, suggesting the multidrug resistance could be stably inherited. Transcriptional measurement of drug target genes and drug transporter genes and deletion analysis of the efflux pump gene cdr4 in the evolved strains suggest that overexpression of cdr4 played a major role in the resistance mechanisms for azoles and terbinafine in the evolved strains, particularly for 30thK2 and 26thV1, and evolved drug-resistant strains had less intracellular ketoconazole accumulation and less disruption of ergosterol accumulations under ketoconazole stress compared to wild type. Mutations specifically present in evolved drug-resistant strains were identified by genome re-sequencing, and drug susceptibility test of knockout mutants for most of mutated genes suggests that mutations in 16 genes, functionally novel in drug resistance, potentially contribute to multidrug resistance in evolved strains.

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Transcription factor CCG-8 plays a pivotal role in azole adaptive responses of Neurospora crassa by regulating intracellular azole accumulation

Cited by 9 publications

References 43 publications

Systematic Prediction of FFAT Motifs Across Eukaryote Proteomes Identifies Nucleolar and Eisosome Proteins With the Predicted Capacity to Form Bridges to the Endoplasmic Reticulum

Systematic Prediction of FFAT Motifs Across Eukaryote Proteomes Identifies Nucleolar and Eisosome Proteins With the Predicted Capacity to Form Bridges to the Endoplasmic Reticulum

Coordinated Regulation of Membrane Homeostasis and Drug Accumulation by Novel Kinase STK-17 in Response to Antifungal Azole Treatment

Experimental Evolution of Multidrug Resistance in Neurospora crassa under Antifungal Azole Stress

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