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Arabidopsis mutant proteomics assay

ABSTRACT: We had a mutant that might be affected for certain protein translation. Triplicate samples were taken from mutant and wild type, each labeled individually for iTRAQ labels. Plant were taken at early vegetative grwoth state for total protein extraction and fractionation, followed by trypsin digestion and peptide analysis using LC-MS/MS.

INSTRUMENT(S): LTQ Orbitrap

ORGANISM(S): Arabidopsis Thaliana (mouse-ear Cress)

TISSUE(S): Whole Body, Leaf

SUBMITTER: Peng Chen

LAB HEAD: Peng Chen

PROVIDER: PXD009770 | Pride | 2019-11-12

REPOSITORIES: Pride

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Publications

AtTrm5a catalyses 1-methylguanosine and 1-methylinosine formation on tRNAs and is important for vegetative and reproductive growth in Arabidopsis thaliana.

Jin Xiaohuan X Lv Zhengyi Z Gao Junbao J Zhang Rui R Zheng Ting T Yin Ping P Li Dongqin D Peng Liangcai L Cao Xintao X Qin Yan Y Persson Staffan S Zheng Bo B Chen Peng P

Nucleic acids research 20190101 2

Modified nucleosides on tRNA are critical for decoding processes and protein translation. tRNAs can be modified through 1-methylguanosine (m1G) on position 37; a function mediated by Trm5 homologs. We show that AtTRM5a (At3g56120) is a Trm5 ortholog in Arabidopsis thaliana. AtTrm5a is localized to the nucleus and its function for m1G and m1I methylation was confirmed by mutant analysis, yeast complementation, m1G nucleoside level on single tRNA, and tRNA in vitro methylation. Arabidopsis attrm5a ...[more]

PMID: 30508117

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Project description:In Hawaii, a rapidly-evolving mutation in the field cricket Teleogryllus oceanicus silences males by interfering with the development of sound-producing structures on their forewings. The mutation is called flatwing (fw), and it persists because of natural selection imposed by an acoustically-orienting parasitoid. We examined gene expression differences between wild-type and mutant crickets, focusing on juvenile wing buds. We profiled mRNA expression levels using RNA-seq, and characterized the wing bud proteome using quantitative mass spectrometry.

2016-03-31 | PXD002451 | Pride

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iTRAQ analysis of ClpP mutant treated with methyl gallate

Project description:Methyl gallate (MG) is an effective microbicide with great potential usefulness in the integrated management of plant diseases and an important potential drug for clinical application. However, its target remains unknown. This study conducted a transposon sequencing (Tn-seq) under MG treatment in plant pathogenic bacterium Ralstonia solanacearum. Tn-seq identified that the mutation of caseinolytic protease proteolytic subunit gene clpP significantly increased the resistance of R. solanacearum to MG, which was validated by the in-frame gene deletion. iTRAQ proteomics analysis was then employed and revealed that chemotaxis and flagella associated proteins were the major substrates degraded by ClpP under the tested condition. Moreover, sulfur metabolism associated proteins were also possible substrates of ClpP and were up-regulated by MG treatment in the wild type R. solanacearum but not in the clpP mutant. Furthermore, molecular docking confirmed the possible interaction between MG and ClpP. Taken together, this study revealed that MG may target bacterial ClpP, inhibit the activity of ClpP, and consequently disorder bacterial proteostasis, providing a theoretical basis for the application of MG.

2020-12-10 | PXD021102 | Pride

PfCDPK7 signaling regulates phospholipid biogenesis in Plasmodium falciparum

Project description:Calcium Dependent Protein Kinases (CDPKs) play important function in calcium signaling at various stage of Plasmodium falciparum. CDPK7 is one such kinase, which plays a crucial role in the development of Plasmodium falciparum parasite. In order to understand the function of PfCDPK7, we performed comparative phosphoproteomic analysis of PfCDPK7 knockout mutant and wild-type lines. In brief, proteins from PfCDPK7 knockout mutant and wild-type parasites were isolated, digested with trypsin and labelled with iTRAQ tags. Phosphopeptides were then enriched from iTRAQ labeled peptides and subjected to LC-MS/MS analysis. This led to the identification of putative substrates of PfCDPK7 in the parasites. A comparative proteomic analysis was performed to measure the protein abundance changes in the PfCDPK7 mutant parasites.

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Functional analysis of HSFB transcription factors [32°C]

Project description:To identify biological function of HSFB transcriptional repressor, we subjected hsfb1 hsfb2b double mutant and wild-type Arabidopsis plant under heat condition (32 Celsius degree) and did microarray experiment. The microarray experiment under non-heated condition is described in GSE14702. We found expression of many heat-shock proteins (HSPs) is not fully enhanced in the double mutant when compared to wild-type plant. This data suggest that the inhibition of expression of HSPs in non-heated condition is important for fully heat-stress response in plant. hsfb1 hsfb2b double mutant vs. wild-type seedlings of Arabidopsis thaliana subjected under heat condition (32 Celsius degree). Biological replicates: 4 replicates.

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Functional analysis of Arabidopsis HSFB transcription factor

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2011-09-06 | E-GEOD-31888 | biostudies-arrayexpress

The landscape of phospho-tyrosine mediated protein interactions of human receptor tyrosine kinases

Project description:Systematically profiling the interactome of 1,144 unique wild-type and mutant phosphotyrosine residues of receptor tyrosine kinases by affinity enrichment - mass spectrometry (AE-MS), followed by a subset of novel interactors validation by competition assay.

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AP-MS analysis of murine and human norovirus VPg interactomes

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