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Quantitative proteomics reveals regulation of KPNA2 and its potential novel cargo proteins in non‐small cell lung cancer
Author(s) -
YU CHIAJUNG,
Wang ChunI,
Chien KunYi,
Wang ChihLiang,
Liu HaoPing,
Cheng ChiaChen,
Chang YuSun,
Yu JauSong
Publication year - 2013
Publication title -
the faseb journal
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.709
H-Index - 277
eISSN - 1530-6860
pISSN - 0892-6638
DOI - 10.1096/fasebj.27.1_supplement.812.1
Subject(s) - biology , gene knockdown , microbiology and biotechnology , e2f1 , stable isotope labeling by amino acids in cell culture , immunoprecipitation , quantitative proteomics , karyopherin , proteomics , proteome , cell cycle , effector , cell , cell culture , nuclear transport , bioinformatics , biochemistry , cell nucleus , gene , genetics , nucleus
The process of nucleocytoplasmic shuttling is mediated by karyopherins. Dysregulated expression of karyopherins may trigger oncogenesis through aberrant distribution of cargo proteins. Previously, we identified karyopherin subunit alpha‐2 (KPNA2) as a potential biomarker for non‐small cell lung cancer (NSCLC). Herein, we applied gene knockdown, subcellular fractionation and SILAC‐based quantitative proteomic strategies to systematically analyze the KPNA2‐regulating protein profiles in NSCLC cell line. Interaction network analysis revealed that KPNA2 involved in signaling for cell cycle, cellular component movements and cell migration. Importantly, E2F1 was identified as a potential novel cargo of KPNA2 in the nuclear proteome. The mRNA levels of potential effectors of E2F1 measured using quantitative PCR indicated that E2F1 is one of the “master molecule” responses to KPNA2 knockdown. Immunofluorescence staining and immunoprecipitation assays disclosed co‐localization and association between E2F1 and KPNA2. An in vitro protein binding assay further demonstrated that E2F1 interacts directly with KPNA2. Moreover, knockdown of KPNA2 led to subcellular redistribution of E2F1 in lung cancer cells. Our results collectively demonstrate the utility of quantitative proteomic approaches and provide a fundamental platform to further explore the biological roles of KPNA2 in NSCLC.