Temporal quantitative proteomics by iTRAQ 2D-LC-MS/MS and corresponding mRNA expression analysis identify post-transcriptional modulation of actin-cytoskeleton regulators during TGF-beta-Induced epithelial-mesenchymal transition.
To gain insights into how TGF-beta 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 cofilin1 potentiated TGF-beta-induced EMT. Further analysis of cofilin1 and beta-actin revealed an increase in their mRNA stability in response to TGF-beta, 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 cofilin1 during TGF-beta-induced EMT.
Keshamouni
Venkateshwar G
VG
Divisions of Pulmonary and Critical Care Medicine, University of Michigan, Ann Arbor, Michigan, 48109, USA. vkeshamo@med.umich.edu
Jagtap
Pratik
P
Michailidis
George
G
Strahler
John R
JR
Kuick
Rork
R
Reka
Ajaya Kumar
AK
Papoulias
Panagiotis
P
Krishnapuram
Rashmi
R
Srirangam
Anjaiah
A
Standiford
Theodore J
TJ
Andrews
Philip C
PC
Omenn
Gilbert S
GS
D000199:Actins / Q000378:metabolism*; D002454:Cell Differentiation / Q000187:drug effects; D045744:Cell Line, Tumor; D002853:Chromatography, Liquid / Q000379:methods*; D051338:Cofilin 1 / Q000378:metabolism; D003599:Cytoskeleton / Q000378:metabolism*; D004848:Epithelium / Q000187:drug effects*; D005786:Gene Expression Regulation; D006801:Humans; D013058:Mass Spectrometry / Q000379:methods*; D008648:Mesoderm / Q000187:drug effects*; D040901:Proteomics / Q000379:methods*; D012323:RNA Processing, Post-Transcriptional; D012333:RNA, Messenger / Q000378:metabolism*; D013997:Time Factors; D016212:Transforming Growth Factor beta / Q000378:metabolism*
D000199:Actins; D051338:Cofilin 1; D012333:RNA, Messenger; D016212:Transforming Growth Factor beta