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A single transcriptional factor Csa3b regulates CRISPR-Cas adaptation and interference in Sulfolobus

ABSTRACT: 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.

ORGANISM(S): Sulfolobus islandicus REY15A

PROVIDER: GSE125156 | GEO | 2019/01/17

REPOSITORIES: GEO

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Coupling transcriptional activation of CRISPR-Cas and DNA repair systems by Csa3a in Sulfolobus islandicus

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.

2017-10-31 | GSE99099 | GEO

RNA-seq data for Sulfolobus islandicus REY15A Δcbp1 strain and the parental strain E234

Project description:RNA-seq data for Sulfolobus islandicus REY15A Δcbp1 strain and the parental strain E234

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A truncated MRE11 form induced by SPRTN is deficient in DNA repair and radiosensitises cancer cells

Project description:he human MRE11/RAD50/NBS1 (MRN) complex plays a crucial role in sensing and repairing DNA DSB. MRE11 possesses 3’-5’ exonuclease and endonuclease activity and forms the core of the multifunctional MRN complex. We previously identified a C-terminally truncated form of MRE11 (TR-MRE11) associated with post-translational MRE11 degradation. Here we identified the approximate cleavage site of TR-MRE11 at 559-580 amino acids, its DNA damage repair function and the factors regulating TR-MRE11 accumulation. The nuclease enzymatic activity of TR-MRE11 was dramatically reduced, associated with a lack of DNA binding efficiency, whilst TR-MRE11 still interacted efficiently with RAD50 and NBS1. Lack of the MRE11 C-terminal resulted in deficient HR repair and increased cellular radiosensitivity. Knockdown of SprT-like N-terminal domain (SPRTN), an essential metalloprotease for DNA-protein crosslink repair, resulted in failure of MRE11 cleavage, with TR-MRE11 protein levels being positively correlated with SPRTN protein expression. The presence of this DNA repair-defective C-terminal truncation could explain the finding of high MRE11 expression, by immunohistochemistry using an antibody against MRE11 prior to the C-terminal, being associated with survival following radical radiotherapy in cancer patients. Ultimately, understanding the functional differences between intact and repair-defective MRE11 may lead to improvements in patient outcomes through a more informed choice of treatment.

2021-02-23 | PXD017964 | Pride

Thermal proteomics of Sulfolobus Islandicus

Project description:Quantitative proteomics of Sulfolobus islandicus after treatment at different temperatures

2021-12-06 | PXD030196 | iProX

Continuous CRISPR spacer acquisition in STSV2-infected crenarchaeon Sulfolobus islandicus REY15A over a 6-day period

Project description:A whole transcriptome study was performed on Sulfolobus islandicus REY15A actively undergoing CRISPR spacer acquisition from the crenarchaeal monocaudavirus STSV2 in rich (TYS) and basal (SCV) media over a 6 day period. Spacer acquisition preceded strong host growth retardation, and changes in viral transcript abundance and virus copy numbers showed significant differences between the two media. Results showed that rich medium favoured CRISPR-Cas immunity generation.

2015-12-06 | E-MTAB-4027 | biostudies-arrayexpress

Sulfolobus islandicus REY15A

Project description:Coupling transcriptional activation of CRISPR-Cas and DNA repair systems by Csa3a in Sulfolobus islandicus

| PRJNA387291 | ENA

Expression data from wild type, mre11delta, rad50delta, and spo11Y135F at meiosis 0 hr and 4 hr.

Project description:Mre11, together with Rad50 and Xrs2/NBS, plays pivotal roles in homologous recombination, repair of DNA double strand breaks (DSBs), activation of damage-induced checkpoint, and telomere maintenance. Using DNA microarray assays to analyze yeast mutants (mre11delta, rad50delta, and spo11Y135F) defective for meiotic DSB formation, we demonstrate that the absence of Mre11 in yeast causes specific effects on regulation of a class of meiotic genes for spore development. The transcriptional deficiency was not observed in other DSB mutants such as rad50delta and spo11Y135F, suggesting the transcriptional defect in mre11delta is due to neither lack of meiotic DSB formation, nor disintegrity of Mre11-Rad50-Xrs2 complex.These defects were confirmed by northern and lacZ reporter gene assays. Keywords: mutant vs. wildtype strains

2007-04-10 | GSE6620 | GEO

A Rad50-null mutation in mouse germ cells causes reduced DSB formation, abnormal DSB end resection and complete loss of germ cells

Project description:Programed DNA double-strand breaks (DSBs) catalyzed by the topoisomerase II-like enzymes, SPO11 and TOPVIBL, initiate meiotic recombination. Following DSB formation, the MRE11-RAD50-NBS1/Xrs2 (MRN/X) complex, along with EXO1 and DNA2, cleave the SPO11-DNA to generate 3′ single-stranded DNA (ssDNA) ends, which are prerequisite for meiotic DSB repair. In both yeast and mammals, MRE11 exhibits endonucleolytic cleavage of the 5′ terminated DNA strand in the vicinity of the DSBs and exonucleolytic resection from 3′ to 5′ towards the DSB ends. RAD50 is a structure maintenance of chromosome (SMC) related protein that contains one ATPase domain at its N- and C- terminal ends, respectively, Zn hook, and anti-parallel coiled coils. RAD50 plays a crucial role in facilitating the MRE11 nuclease activity on DSBs by ATP binding and hydrolysis. However, the in vivo function of Rad50 in mammalian germ cells, particularly its in vivo role in the resection of meiotic DSB ends at the molecular level remains elusive. Here, we performed END-seq with synchronized zygotene spermatocytes from control and germ cell specific Rad50 mutant (Rad50-sKO) mice. We find that the number of formed DSB in the mutant spermatocytes was reduced compared to control spermatocytes (6 636 DSBs in Rad50-sKO spermatocytes vs 8 168 in control spermatocytes) and abnormal DSB end resection occurred in mutant spermatocytes (DSB end resection length: 1 279 nts in Rad50-sKO spermatocytes vs 1 923 nts in control spermatocytes). Thus, RAD50 is essential for DSB formation and end resection during mammalian meiosis.

2024-04-02 | GSE262706 | GEO

Transcription profiling of S. cerevisiae mre11, rad50 and spo11 mutants in premeiosis and meiotic prophase

2007-10-23 | E-GEOD-6620 | biostudies-arrayexpress

Sulfolobus islandicus HCD+ETD, including LFQ

Project description:Protein posttranslational methylation and acetylations have been reported to occur in archaea, including members of the genus Sulfolobus, but have not been characterized on a proteome-wide scale. Sulfolobus chromatin proteins are known to be methylated and acetylated on lysine side chains, resembling eukaryotic histones in this respect. We utilized bottom-up and top-down proteomic approaches to perform a global and deep methylation study in the hyperthermoacidophylic archaeon S. islandicus LAL 14/1 with a particular focus on chromatin proteins. Without specific enrichment, 1931 methylation sites on 731 protein were found by bottom-up proteomic analysis. The dynamics of protein methylation has been investigated on 424-526 proteins throughout 3 cell culture growth stages. We also confirm the relaxed specificity and steady abundance of the previously described methyltransferase aKMT4 which is implicated in the massive proteome methylation. We also detected an abundant N-terminal acetylation of S. islandicus proteins, with more than one third of the detected N-terminal peptides being acetylated

2022-03-01 | PXD003102 | Pride

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