A single transcriptional factor Csa3b regulates CRISPR-Cas adaptation and interference in Sulfolobus
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ABSTRACT:
PROVIDER: PRJNA515414 | ENA |
REPOSITORIES: ENA
Dataset's files
| Action | DRS | |||
|---|---|---|---|---|
| SRR8447230_1.fastq.gz | Fastqsanger.gz | |||
| SRR8447230_2.fastq.gz | Fastqsanger.gz | |||
| SRR8447227_1.fastq.gz | Fastqsanger.gz | |||
| SRR8447227_2.fastq.gz | Fastqsanger.gz | |||
| SRR8447228_1.fastq.gz | Fastqsanger.gz |
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Project description:Transcriptome sequencing was carried out on an Illumina HiSeq platform to investigate CRISPR-Cas and DNA repair systems by Csa3b in Sulfolobus islandicus Rey15A. We compared the differently expressed genes in Sulfolobus islandicus Rey15A strain with csa3a overexpression vs. Sulfolobus islandicus Rey15A strain carrying an empty expression vector,We find thatcmr-α (SiRe_0890 ~ SiRe_0895) and cmr-β (SiRe_0597 ~ SiRe_0603)、the DNA double strand break (DSB)repair genes, including nurA, rad50, mre11, and herA (SiRe_0061 ~ SiRe_0064), as well as two subunits of DNA polymerase II (SiRe_0615 and SiRe_0617) that function in DNA repair, were significantly up-regulated. Our data indicated that the Csa3b regulator couples transcriptional activation of cmr genes, DNA repair genes.
2019-01-17 | GSE125156 | GEO
Project description:Transcriptome sequencing was carried out on an Illumina HiSeq platform to investigate the activation of CRISPR-Cas and DNA repair systems by Csa3a in Sulfolobus islandicus Rey15A. We compared the differently expressed genes in Sulfolobus islandicus Rey15A strain with csa3a overexpression vs. Sulfolobus islandicus Rey15A strain carrying an empty expression vector, cas1 deletion strain with csa3a overexpression vs. cas1 deletion strain carrying an empty expression vector, as well as interference-deficient strain with csa3a overexpression vs. interference-deficient strain carrying an empty expression vector. We find that cas genes (SiRe_0760, SiRe_0761, SiRe_0762, SiRe_0763), nucleotidyltransferase domain of DNA polymerase beta (SiRe_0459), chromosome segregation protein (SMC)-related ATPase (SiRe_0649), SMC-related protein (SiRe_1142) and three HerA helicases involved in DNA double break repair (encoded by SiRe_0064 and SiRe_0095 of nurA-herA operons, and SiRe_1857) were significantly up-regulated. Our data indicated that the Csa3a regulator couples transcriptional activation of spacer acquisition genes, CRISPR RNA transcription, DNA repair and genome stability genes.
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Project description:Bacteria protect themselves from infection by bacteriophages (phages) using different defence systems, such as CRISPR-Cas. Although CRISPR-Cas provides phage resistance, fitness costs are incurred, such as through autoimmunity. CRISPR-Cas regulation can optimise defence and minimise these costs. We recently developed a genome-wide functional genomics approach (SorTn-seq) for high-throughput discovery of regulators of bacterial gene expression. Here, we applied SorTn-seq to identify loci influencing expression of the two type III-A Serratia CRISPR arrays. Multiple genes affected CRISPR expression, including those involved in outer membrane and lipopolysaccharide synthesis. By comparing loci affecting type III CRISPR arrays and cas operon expression, we identified PigU (LrhA) as a repressor that co-ordinately controls both arrays and cas genes. By repressing type III-A CRISPR-Cas expression, PigU shuts off CRISPR-Cas interference against plasmids and phages. PigU also represses interference and CRISPR adaptation by the type I-F system, which is also present in Serratia. RNA sequencing demonstrated that PigU is a global regulator that controls secondary metabolite production and motility, in addition to CRISPR-Cas immunity. Increased PigU also resulted in elevated expression of three Serratia prophages, indicating their likely induction upon sensing PigU-induced cellular changes. In summary, PigU is a major regulator of CRISPR-Cas immunity in Serratia.
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