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The genome-scale binding profile of the CCAAT-binding complex in Aspergillus fumigatus

ABSTRACT: The genome-scale binding profile of the CCAAT-binding complex in Aspergillus fumigatus

PROVIDER: PRJNA533301 | ENA |

REPOSITORIES: ENA

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The genome-scale binding profile of the CCAAT-binding complex in Aspergillus fumigatus

Project description:The CCAAT-binding complex (CBC) is a heterotrimetric transcription factor conserved in all eukaryotes. The CBC specifically interacts with the pentanucleotide motif known as the CCAAT box, which is one of the most ubiquitous cis-acting elements on the eukaryotic promoters. In Aspergillus fumigatus, the CBC consists of three subunits HapB, HapC, and HapE, which are all required for specific binding to the target sequences. This regulatory complex has been shown to be involved in the regulation of a wide range of cellular processes, including primary and secondary metabolism, iron homeostasis, as well as azole drug resistance and virulence in mammalian hosts. In this study, we investigated the genome-wide binding profile of the CBC using the chromatin immunoprecipitation followed by high-throughput sequencing (ChIP-seq). We identified over 1,000 peak regions, which showed more than 5-fold enrichment compared to the input control in steady-state iron-replete (+Fe) and iron-starvation (-Fe) conditions. Bioinformatic analysis of the peak regions revealed the 5’-CSAATVR-3’ sequence as the most enriched nucleotide motif, suggesting that this motif acts as the consensus binding sequence for the CBC in vivo.

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The CCAAT-Binding Complex Controls Mitochondrial Activity and Iron Homeostasis in Candida glabrata

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The Negative Cofactor 2 complex is a master regulator of drug resistance in Aspergillus fumigatus [ChIP-seq]

Project description:The Negative cofactor 2 (NCT) complex is an evolutionally conserved heterodimeric transcription factor. In Aspergillus fumigatus, the NCT complex consists of two subunits NctA and NctB. Through a genome-wide screening of a transcription factor null mutant strains, we found that loss of the NCT complex leads to a multi-drug resistance phenotype including the azoles (itraconazole, voriconazole and posaconazole) as well as the salvage therapeutic amphotericin B, and terbinafine. To obtain further insight into the molecular mechanisms driving the azole-resistance in the NCT complex null mutants, we analyzed genome-wide binding profiles of NctA using chromatin-immunoprecipitation sequencing (ChIP-seq). Our ChIP-seq analysis revealed that NCT complex binds the promoters of several ergosterol biosynthetic genes, their transcriptional regulators, and the azole efflux pump cdr1B. Taken together, these results suggest that the NCT complex plays a role as a master regulator of drug resistance in A. fumigatus.

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The fungal CCAAT-binding complex and HapX display highly variable but evolutionary conserved synergetic promoter-specific DNA recognition

Project description:To sustain iron homeostasis, microorganisms have evolved fine-tuned mechanisms for uptake, storage and detoxification of the essential metal iron. In the human pathogen Aspergillus fumigatus, the fungal-specific bZIP-type transcription factor HapX coordinates adaption to both iron starvation and iron excess and is thereby crucial for virulence. Previous studies indicated that a HapX homodimer interacts with the CCAAT-binding complex (CBC) to cooperatively bind bipartite DNA motifs; however, the significance of the HapX and CBC interaction and the mode of HapX-DNA recognition had not been resolved. In this study, we characterized the genome-wide binding profiles of HapX using the chromatin immunoprecipitation followed by high-throughput sequencing (ChIP-seq). Our ChIP-seq results, in combination with in vitro surface plasmon resonance analysis, phylogenetic comparison, and genetic analysis, revealed an astonishing plasticity of the CBC:HapX DNA-recognition mode.

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