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Screening of Candida albicans GRACE library revealed a unique pattern of biofilm construction under repression of the essential gene ILS1

ABSTRACT: Screening of Candida albicans GRACE library revealed a unique pattern of biofilm construction under repression of the essential gene ILS1

PROVIDER: PRJNA525426 | ENA |

REPOSITORIES: ENA

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			Action	DRS
	SRR8664500_1.fastq.gz	Fastqsanger.gz
	SRR8664500_2.fastq.gz	Fastqsanger.gz
	SRR8664495_1.fastq.gz	Fastqsanger.gz
	SRR8664495_2.fastq.gz	Fastqsanger.gz
	SRR8664496_1.fastq.gz	Fastqsanger.gz

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Screening of Candida albicans GRACE library revealed a unique pattern of biofilm construction under repression of the essential gene ILS1

Project description:ILS1 mutated Candida albicans was unable to grow as yeast-phase cells but was capable of producing a tridimensional biofilm structure in spite of reduced metabolic activity. This biofilm still relied on the classical biofilm genes, while it differentially induced groups of genes involved in adhesion, protein synthesis, cell wall organization, and protein folding. Although the conditional mutant repressed genes annotated for morphology and homeostasis processes affecting morphology and metabolism, the dynamic cell growth enabled the construction of a complex biofilm community independent of ILS1.

2019-03-08 | GSE127796 | GEO

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Project description:The incidence of candidiasis, caused mainly by the opportunistic fungal Candida albicans, remains a major concern in public health due to the emerging antifungal resistance. C. albicans modulates their proteome under different conditions trying to cope with several stressors in host niches. In this study we developed a C. albicans spectral library by analysing a total of 52 samples corresponding to cells treated with 5 and 10 mM hydrogen peroxide, 40 and 60 mM acetic acid and in the interaction with human macrophages (THP-1) at 3, 6 and 9 h (4 biological replicates). This samples represented several morphologies of C. albicans (yeast and hypha) and different culture conditions (temperature, media…) that contributed to the incorporation of different proteins into the library. The analysis of these conditions by DDA mode allowed the construction of a C. albicans spectral library comprising information of 2879 proteins and 25645 peptides which entailed a 46.5% of the total proteome. The C. albicans spectral library created constitutes a powerful proteomic tool available for the scientific community and could be useful for global (DIA) or targeted (SRM and PRM) studies.

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Project description:Next-generation sequencing has been widely used for the genome-wide profiling of histone modifications, transcription factor binding and gene expression through chromatin immunoprecipitated DNA sequencing (ChIP-seq) and cDNA sequencing (RNA-seq). Here, we describe a versatile library construction method that can be applied to both ChIP-seq and RNA-seq on the widely used Illumina platforms. Standard methods for ChIP-seq library construction require nanograms of starting DNA, substantially limiting its application to rare cell types or limited clinical samples. By minimizing the DNA purification steps that cause major sample loss, our method achieved a high sensitivity in ChIP-seq library preparation. Using this method, we achieved the following: (1) generated high-quality epigenomic and transcription factor-binding maps using ChIP-seq for murine adipocytes; (2) successfully prepared a ChIP-seq library from as little as 25 pg of starting DNA; (3) achieved paired-end sequencing of the ChIP-seq libraries; (4) systematically profiled gene expression dynamics during murine adipogenesis using RNA-seq; and (5) preserved the strand specificity of the transcripts in RNA-seq. Given its sensitivity and versatility in both double-stranded and single-stranded DNA library construction, this method has wide applications in genomic, epigenomic, transcriptomic and interactomic studies.

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Screening of Candida albicans GRACE library revealed a unique pattern of biofilm formation under repression of the essential gene ILS1.

Project description:Candida albicans biofilm formation is governed by a regulatory circuit comprising nine transcription factors which control a network of target genes. However, there are still unknown genes contributing to biofilm features. Thus, the GRACE library was screened to identify genes involved in mature biofilm development. Twenty-nine conditional mutants were selected for a second screening revealing three groups of genes: twenty- two conditional mutants were defective for normal growth and unable to form biofilms; six strains, conditionally defective in genes ARC40, ARC35, ORF19.2438, SKP1, ERG6, and ADE5,7 that are likely essential or involved in general cell processes, grew normally as free-floating cells but produced less biofilm; finally, the conditional strain for a putative essential isoleucyl- tRNA synthetase gene, ILS1, was unable to grow as yeast-phase cells but was capable of producing a tridimensional biofilm structure in spite of reduced metabolic activity. This unique biofilm still relied on the classical biofilm genes, while it differentially induced groups of genes involved in adhesion, protein synthesis, cell wall organization, and protein folding. Although the conditional mutant repressed genes annotated for morphology and homeostasis processes affecting morphology and metabolism, the dynamic cell growth enabled the formation of a complex biofilm community independent of ILS1.

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Project description:Background: Next-generation sequencing (NGS) is fundamental to the current biological and biomedical research. Construction of sequencing library is a key step of NGS. Therefore, various library construction methods have been explored. However, the current methods are still limited by some shortcomings. Results: This study developed a new NGS library construction method, Single strand Adaptor Library Preparation (SALP), by using a novel single strand adaptor (SSA). SSA is a double-stranded oligonucleotide with a 3ʹ overhang of 3 random nucleotides, which can be efficiently ligated to the 3′ end of single strand DNA by T4 DNA ligase. SALP can be started with any denatured DNA fragments such as those sheared by Tn5 tagmentation, enzyme digestion and sonication. When started with Tn5-tagmented chromatin, SALP can overcome a key limitation of ATAC-seq and become a high-throughput NGS library construction method, SALP-seq, which can be used to comparatively characterize the chromatin openness state of multiple cells unbiasly. In this way, this study successfully characterized the comparative chromatin openness states of four different cell lines, including GM12878, HepG2, HeLa and 293T, with SALP-seq. Similarly, this study also successfully characterized the chromatin openness states of HepG2 cells with SALP-seq by using 105 to 500 cells. Conclusions: This study developed a new NGS library construction method, SALP, by using a novel kind of single strand adaptor (SSA), which should has wide applications in the future due to its unique performance.

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Reduction of rRNA during sRNA library construction using blocking oligos

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Gene expression pattern of IFU5 deleted Candida albicans cells

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