Identification of the −1 translational frameshift sites using a liquid chromatography–tandem mass spectrometric approach

Tsay, Yeou Guang; Chen, Chang Chieh; Hu, Shiau Ting

doi:10.1016/j.ab.2004.12.031

Cited by 6 publications

(5 citation statements)

References 37 publications

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“…The 17 kDa protein fraction from gel-filtration column chromatography fractionation was further separated by 12% SDS-PAGE with Coomassie blue staining. The gel of 17 kDa position was cut and then soaked in 25 mM NH 4 HCO 3 for 10 min, and then incubated in 25 mM NH 4 HCO 3 containing 50% acetonitrile for another 10 min, and then was subjected to in-gel tryptic digestion as descripted by Tsay et al [ 35 ]. After digestion, the samples were purified and subjected to LC/MS/MS analysis.…”

Section: Methodsmentioning

confidence: 99%

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Multiple Patterns of Regulation and Overexpression of a Ribonuclease-Like Pathogenesis-Related Protein Gene, OsPR10a, Conferring Disease Resistance in Rice and Arabidopsis

Huang

Lin

et al. 2016

PLoS ONE

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An abundant 17 kDa RNase, encoded by OsPR10a (also known as PBZ1), was purified from Pi-starved rice suspension-cultured cells. Biochemical analysis showed that the range of optimal temperature for its RNase activity was 40–70°C and the optimum pH was 5.0. Disulfide bond formation and divalent metal ion Mg2+ were required for the RNase activity. The expression of OsPR10a::GUS in transgenic rice was induced upon phosphate (Pi) starvation, wounding, infection by the pathogen Xanthomonas oryzae pv. oryzae (Xoo), leaf senescence, anther, style, the style-ovary junction, germinating embryo and shoot. We also provide first evidence in whole-plant system, demonstrated that OsPR10a-overexpressing in rice and Arabidopsis conferred significant level of enhanced resistance to infection by the pathogen Xoo and Xanthomona campestris pv. campestris (Xcc), respectively. Transgenic rice and Arabidopsis overexpressing OsPR10a significantly increased the length of primary root under phosphate deficiency (-Pi) condition. These results showed that OsPR10a might play multiple roles in phosphate recycling in phosphate-starved cells and senescing leaves, and could improve resistance to pathogen infection and/or against chewing insect pests. It is possible that Pi acquisition or homeostasis is associated with plant disease resistance. Our findings suggest that gene regulation of OsPR10a could act as a good model system to unravel the mechanisms behind the correlation between Pi starvation and plant-pathogen interactions, and also provides a potential application in crops disease resistance.

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Section: Methodsmentioning

confidence: 99%

“…After digestion, the samples were purified and subjected to LC/MS/MS analysis. Amino acid sequence data processing was conducted as described by Tsay et al [ 35 , 36 ].…”

Section: Methodsmentioning

confidence: 99%

Multiple Patterns of Regulation and Overexpression of a Ribonuclease-Like Pathogenesis-Related Protein Gene, OsPR10a, Conferring Disease Resistance in Rice and Arabidopsis

Huang

Lin

et al. 2016

PLoS ONE

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“…GST‐GAP was phosphorylated in vitro with Cdk1 as above, separated by SDS–PAGE and stained with colloidal coomassie blue. The protein in the gel piece was subjected to tryptic digestion as described previously [22]. Briefly, the gel was washed and dried in a Speed‐Vac (Savant), the protein was reduced with β‐mercaptoethanol and then modified with 4‐vinylpyridine.…”

Section: Methodsmentioning

confidence: 99%

Phosphoregulation of MgcRacGAP in mitosis involves Aurora B and Cdk1 protein kinases and the PP2A phosphatase

et al. 2008

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“…Following extracellular chemical cross-linking, the SARS-CoV VLP-pre-incubated Vero E6 cells were lysed with Empigen BB lysis buffer (50 mM Tris-HCl, pH 7.4, 0.05 % sodium deoxycholate, 150 mM NaCl, 1 mM EDTA, and 0.3 % Empigen BB) supplemented with protease inhibitor cocktail (Roche) and phenylmethylsulfonyl fluoride. The protein lysates were subjected to immunoprecipitation followed by sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE), silver staining, and LC-MS/MS analysis as described previously [ 23 , 24 ].…”

Section: Methodsmentioning

confidence: 99%

Surface vimentin is critical for the cell entry of SARS-CoV

et al. 2016

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BackgroundSevere acute respiratory syndrome coronavirus (SARS-CoV) caused a global panic due to its high morbidity and mortality during 2002 and 2003. Soon after the deadly disease outbreak, the angiotensin-converting enzyme 2 (ACE2) was identified as a functional cellular receptor in vitro and in vivo for SARS-CoV spike protein. However, ACE2 solely is not sufficient to allow host cells to become susceptible to SARS-CoV infection, and other host factors may be involved in SARS-CoV spike protein-ACE2 complex.ResultsA host intracellular filamentous cytoskeletal protein vimentin was identified by immunoprecipitation and LC-MS/MS analysis following chemical cross-linking on Vero E6 cells that were pre-incubated with the SARS-CoV spike protein. Moreover, flow cytometry data demonstrated an increase of the cell surface vimentin level by 16.5 % after SARS-CoV permissive Vero E6 cells were treated with SARS-CoV virus-like particles (VLPs). A direct interaction between SARS-CoV spike protein and host surface vimentin was further confirmed by far-Western blotting. In addition, antibody neutralization assay and shRNA knockdown experiments indicated a vital role of vimentin in cell binding and uptake of SARS-CoV VLPs and the viral spike protein.ConclusionsA direct interaction between vimentin and SARS-CoV spike protein during viral entry was observed. Vimentin is a putative anti-viral drug target for preventing/reducing the susceptibility to SARS-CoV infection.

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Identification of the −1 translational frameshift sites using a liquid chromatography–tandem mass spectrometric approach

Cited by 6 publications

References 37 publications

Multiple Patterns of Regulation and Overexpression of a Ribonuclease-Like Pathogenesis-Related Protein Gene, OsPR10a, Conferring Disease Resistance in Rice and Arabidopsis

Multiple Patterns of Regulation and Overexpression of a Ribonuclease-Like Pathogenesis-Related Protein Gene, OsPR10a, Conferring Disease Resistance in Rice and Arabidopsis

Phosphoregulation of MgcRacGAP in mitosis involves Aurora B and Cdk1 protein kinases and the PP2A phosphatase

Surface vimentin is critical for the cell entry of SARS-CoV

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