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)

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

88 Scopus citations

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

This output contributes to the following UN Sustainable Development Goals (SDGs)

Publisher link

10.1021/pr8006478

Find It

Find It at U of M Libraries

Fingerprint

Dive into the research topics of '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'. Together they form a unique fingerprint.

Cite this

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.

@article{40526ec7e1434539923dede5c876f1ad,

title = "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 = "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.",

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

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

year = "2009",

month = jan,

doi = "10.1021/pr8006478",

language = "English (US)",

volume = "8",

pages = "35--47",

journal = "Journal of Proteome Research",

issn = "1535-3893",

publisher = "American Chemical Society",

number = "1",

}

TY - JOUR

T1 - 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

AU - Keshamouni, Venkateshwar G.

AU - Jagtap, Pratik

AU - Michailidis, George

AU - Strahler, John R.

AU - Kuick, Rork

AU - Reka, Ajaya Kumar

AU - Papoulias, Panagiotis

AU - Krishnapuram, Rashml

AU - Srirangam, Anjaiah

AU - Standiford, Theodore J.

AU - Andrews, Philip C.

AU - Omenn, Gilbert S.

PY - 2009/1

Y1 - 2009/1

N2 - 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.

AB - 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.

KW - Actin-cytoskeleton remodeling

KW - Beta-actin

KW - Calpain

KW - Cofilin1

KW - Epithelial-mesenchymal transition

KW - Ezrin

KW - ITRAQ

KW - Lung cancer

KW - Moesin

KW - Qualitative proteomics

KW - TGF-β

UR - https://www.scopus.com/pages/publications/60849087975

UR - https://www.scopus.com/pages/publications/60849087975#tab=citedBy

U2 - 10.1021/pr8006478

DO - 10.1021/pr8006478

M3 - Article

C2 - 19118450

AN - SCOPUS:60849087975

SN - 1535-3893

VL - 8

SP - 35

EP - 47

JO - Journal of Proteome Research

JF - Journal of Proteome Research

IS - 1

ER -

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

Publisher link

Find It

Other files and links

Fingerprint

Cite this