Temporal quantitative proteomics by iTRAQ 2D-LC-MS/MS and corresponding mRNA expression analysis identify post-transcriptional modulation of actin-cytoskeleton regulators during TGF-β-Lnduced epithelial-mesenchymal transition

Venkateshwar G. Keshamouni; Pratik Jagtap; George Michailidis; John R. Strahler; Rork Kuick; Ajaya Kumar Reka; Panagiotis Papoulias; Rashml Krishnapuram; Anjaiah Srirangam; Theodore J. Standiford; Philip C. Andrews; Gilbert S. Omenn

doi:10.1021/pr8006478

Temporal quantitative proteomics by iTRAQ 2D-LC-MS/MS and corresponding mRNA expression analysis identify post-transcriptional modulation of actin-cytoskeleton regulators during TGF-β-Lnduced epithelial-mesenchymal transition

Venkateshwar G. Keshamouni, Pratik Jagtap, George Michailidis, John R. Strahler, Rork Kuick, Ajaya Kumar Reka, Panagiotis Papoulias, Rashml Krishnapuram, Anjaiah Srirangam, Theodore J. Standiford, Philip C. Andrews, Gilbert S. Omenn

Biochemistry, Molecular Biology, and Biophysics (TMED)

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Abstract

To gain insights into how TGF-β regulates epithelial-mesenchymal transition (EMT), we assessed the time course of proteins and mRNAs during EMT by multiplex iTRAQ labeling and 2D-LC-MS/MS, and by hybridization, respectively. Temporal iTRAQ analysis identified 66 proteins as differentially expressed during EMT, including newly associated proteins calpain, fascin and macrophage-migration inhibitory factor (MIF). Comparing protein and mRNA expression overtime showed that all the 14 up-regulated proteins involved in the actin-cytoskeleton remodeling were accompanied by increases in corresponding mRNA expression. Interestingly, siRNA mediated knockdown of cofilinl potentiated TGFβ-induced EMT. Further analysis of cofilinl and β-actin revealed an increase in their mRNA stability in response to TGF-β, contributing to the observed increase in mRNA and protein expression. These results are the first demonstration of post-transcriptional regulation of cytoskeletal remodelling and a key role for cofilinl during TGF-β-induced EMT.

Original language	English (US)
Pages (from-to)	35-47
Number of pages	13
Journal	Journal of Proteome Research
Volume	8
Issue number	1
DOIs	https://doi.org/10.1021/pr8006478
State	Published - Jan 2009

Keywords

Actin-cytoskeleton remodeling
Beta-actin
Calpain
Cofilin1
Epithelial-mesenchymal transition
Ezrin
ITRAQ
Lung cancer
Moesin
Qualitative proteomics
TGF-β

UN SDGs

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Keshamouni, V. G., Jagtap, P., Michailidis, G., Strahler, J. R., Kuick, R., Reka, A. K., Papoulias, P., Krishnapuram, R., Srirangam, A., Standiford, T. J., Andrews, P. C., & Omenn, G. S. (2009). Temporal quantitative proteomics by iTRAQ 2D-LC-MS/MS and corresponding mRNA expression analysis identify post-transcriptional modulation of actin-cytoskeleton regulators during TGF-β-Lnduced epithelial-mesenchymal transition. Journal of Proteome Research, 8(1), 35-47. https://doi.org/10.1021/pr8006478

Temporal quantitative proteomics by iTRAQ 2D-LC-MS/MS and corresponding mRNA expression analysis identify post-transcriptional modulation of actin-cytoskeleton regulators during TGF-β-Lnduced epithelial-mesenchymal transition. / Keshamouni, Venkateshwar G.; Jagtap, Pratik; Michailidis, George et al.
In: Journal of Proteome Research, Vol. 8, No. 1, 01.2009, p. 35-47.

Research output: Contribution to journal › Article › peer-review

Keshamouni, VG, Jagtap, P, Michailidis, G, Strahler, JR, Kuick, R, Reka, AK, Papoulias, P, Krishnapuram, R, Srirangam, A, Standiford, TJ, Andrews, PC & Omenn, GS 2009, 'Temporal quantitative proteomics by iTRAQ 2D-LC-MS/MS and corresponding mRNA expression analysis identify post-transcriptional modulation of actin-cytoskeleton regulators during TGF-β-Lnduced epithelial-mesenchymal transition', Journal of Proteome Research, vol. 8, no. 1, pp. 35-47. https://doi.org/10.1021/pr8006478

Keshamouni VG, Jagtap P, Michailidis G, Strahler JR, Kuick R, Reka AK et al. Temporal quantitative proteomics by iTRAQ 2D-LC-MS/MS and corresponding mRNA expression analysis identify post-transcriptional modulation of actin-cytoskeleton regulators during TGF-β-Lnduced epithelial-mesenchymal transition. Journal of Proteome Research. 2009 Jan;8(1):35-47. doi: 10.1021/pr8006478

Keshamouni, Venkateshwar G. ; Jagtap, Pratik ; Michailidis, George et al. / Temporal quantitative proteomics by iTRAQ 2D-LC-MS/MS and corresponding mRNA expression analysis identify post-transcriptional modulation of actin-cytoskeleton regulators during TGF-β-Lnduced epithelial-mesenchymal transition. In: Journal of Proteome Research. 2009 ; Vol. 8, No. 1. pp. 35-47.

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abstract = "To gain insights into how TGF-β regulates epithelial-mesenchymal transition (EMT), we assessed the time course of proteins and mRNAs during EMT by multiplex iTRAQ labeling and 2D-LC-MS/MS, and by hybridization, respectively. Temporal iTRAQ analysis identified 66 proteins as differentially expressed during EMT, including newly associated proteins calpain, fascin and macrophage-migration inhibitory factor (MIF). Comparing protein and mRNA expression overtime showed that all the 14 up-regulated proteins involved in the actin-cytoskeleton remodeling were accompanied by increases in corresponding mRNA expression. Interestingly, siRNA mediated knockdown of cofilinl potentiated TGFβ-induced EMT. Further analysis of cofilinl and β-actin revealed an increase in their mRNA stability in response to TGF-β, contributing to the observed increase in mRNA and protein expression. These results are the first demonstration of post-transcriptional regulation of cytoskeletal remodelling and a key role for cofilinl during TGF-β-induced EMT.",

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Temporal quantitative proteomics by iTRAQ 2D-LC-MS/MS and corresponding mRNA expression analysis identify post-transcriptional modulation of actin-cytoskeleton regulators during TGF-β-Lnduced epithelial-mesenchymal transition

Abstract

Keywords

UN SDGs

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