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Sulfolobus islandicus LC-MSMS - aKMT catalyzes extensive protein lysine methylation in the hyperthermophilic archaeon Sulfolobus islandicus but is dispensable for the growth of the organism

ABSTRACT: Analysis of the methylation of total cellular protein by mass spectrometry revealed that methylated proteins accounted for ~2/3 (1,158/1,750) and ~1/3 (591/1,766) of the identified proteins in the wild type and the mutant strains, respectively, indicating that there is extensive protein methylation in S. islandicus and that aKMT is a major protein methyltransferase in this organism.

INSTRUMENT(S): LTQ FT, LTQ Orbitrap

ORGANISM(S): Sulfolobus Islandicus

TISSUE(S): Cell Culture

SUBMITTER: yindi chu

LAB HEAD: Li Huang

PROVIDER: PXD003424 | Pride | 2016-06-28

REPOSITORIES: Pride

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			Action	DRS
	MS140628-M.msf	Msf
	MS140628-M.pep.xml	Pepxml
	MS140628-M.prot.xml	Xml
	MS140628-M1.raw	Raw
	MS140628-M10.raw	Raw

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Publications

aKMT Catalyzes Extensive Protein Lysine Methylation in the Hyperthermophilic Archaeon Sulfolobus islandicus but is Dispensable for the Growth of the Organism.

Chu Yindi Y Zhu Yanping Y Chen Yuling Y Li Wei W Zhang Zhenfeng Z Liu Di D Wang Tongkun T Ma Juncai J Deng Haiteng H Liu Zhi-Jie ZJ Ouyang Songying S Huang Li L

Molecular & cellular proteomics : MCP 20160621 9

Protein methylation is believed to occur extensively in creanarchaea. Recently, aKMT, a highly conserved crenarchaeal protein lysine methyltransferase, was identified and shown to exhibit broad substrate specificity in vitro Here, we have constructed an aKMT deletion mutant of the hyperthermophilic crenarchaeon Sulfolobus islandicus The mutant was viable but showed a moderately slower growth rate than the parental strain under non-optimal growth conditions. Consistent with the moderate effect of ...[more]

PMID: 27329856

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Sulfolobus islandicus HCD+ETD, including LFQ

2022-03-01 | PXD003102 | Pride

Sulfolobus islandicus top-down LC-MSMS

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 with particular interest in chromatin proteins. Without specific enrichment, 731 protein were found by bottom-up proteomic analysis. The methylation sites on >400 proteins were monitored throughout 3 cell culture growth stages: early exponential, late exponential and stationary. (The previously described aKMT4 is found to be a plausible methyltransferase responsible for the massive methylation.) The proteome-wide top-down study/approach revealed 3778 proteoforms of 681 proteins, including 292 methylated proteoforms, of which 85 were comprehensively characterized by high-resolution MS/MS. Methylated proteoforms of the five chromatin proteins were characterized in detail by combination of bottom-up and top-down data showing the differences between the closely related Sul7d proteins. The two Alba chromatin proteins are, for the first time, reported to be methylated in this work. Alba1 protein is shown to be mono-, di- and trimethylated at the lysine-16 side chain. Stage-wise top-down analysis shows that the relative abundance of the methylated proteoforms versus non-methylated ones significantly grows/increases for Alba1 and Cren7 chromatin proteins throughout the cell growth. These findings highlight the ubiquitous lysine methylation throughout the S. islandicus proteome and singificantly enrich our knowledge related aboutarchaeal chromatin proteins.

2016-09-13 | PXD003074 | Pride

Sulfolobus islandicus PTM and quantitative study using bottom-up proteomics

Project description:Protein posttranslational methylation and acetylation have been reported to occur in archaea, including the genus Sulfolobus, but have never been characterized on a proteome-wide scale. Among important Sulfolobus proteins carrying such modifications are the chromatin proteins that have been described to be methylated and acetylated on lysine side chains, resembling eukaryotic histones in that aspect. To get more insight into the extent of these modifications and their dynamics during the different growth steps of the thermoacidophylic archaeon S. islandicus , we performed a global and deep proteomics analysis using a combination of high-throughput bottom-up and top-down proteomics approaches on a single high-resolution mass spectrometer. 1,931 methylation sites on 751 proteins were found by the bottom-up analysis, with methylation sites of 424-526 proteins monitored throughout three cell culture growth stages: Early, Mid and Late. The previously described aKMT4 methyltransferase was found to be potentially responsible for this massive modification.

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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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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.

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Thermal proteomics of Sulfolobus Islandicus

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Functional analysis of the NucS/EndoMS of the hyperthermophilic archaeon Sulfolobus islandicus REY15A

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