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-beta-Induced epithelial-mesenchymal transition.
abstract: 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.
authors.affiliation: Divisions of Pulmonary and Critical Care Medicine, University of Michigan, Ann Arbor, Michigan, 48109, USA. vkeshamo@med.umich.edu
authors.forename: Venkateshwar G
authors.initials: VG
authors.lastname: Keshamouni
authors.forename: Pratik
authors.initials: P
authors.lastname: Jagtap
authors.forename: George
authors.initials: G
authors.lastname: Michailidis
authors.forename: John R
authors.initials: JR
authors.lastname: Strahler
authors.forename: Rork
authors.initials: R
authors.lastname: Kuick
authors.forename: Ajaya Kumar
authors.initials: AK
authors.lastname: Reka
authors.forename: Panagiotis
authors.initials: P
authors.lastname: Papoulias
authors.forename: Rashmi
authors.initials: R
authors.lastname: Krishnapuram
authors.forename: Anjaiah
authors.initials: A
authors.lastname: Srirangam
authors.forename: Theodore J
authors.initials: TJ
authors.lastname: Standiford
authors.forename: Philip C
authors.initials: PC
authors.lastname: Andrews
authors.forename: Gilbert S
authors.initials: GS
authors.lastname: Omenn
mesh_terms: 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*
chemical_list: D000199:Actins; D051338:Cofilin 1; D012333:RNA, Messenger; D016212:Transforming Growth Factor beta