title: A functional siRNA screen identifies genes modulating angiotensin II-mediated EGFR transactivation.
abstract: The angiotensin type 1 receptor (AT1R) transactivates the epidermal growth factor receptor (EGFR) to mediate cellular growth, however, the molecular mechanisms involved have not yet been resolved. To address this, we performed a functional siRNA screen of the human kinome in human mammary epithelial cells that demonstrate a robust AT1R-EGFR transactivation. We identified a suite of genes encoding proteins that both positively and negatively regulate AT1R-EGFR transactivation. Many candidates are components of EGFR signalling networks, whereas others, including TRIO, BMX and CHKA, have not been previously linked to EGFR transactivation. Individual knockdown of TRIO, BMX or CHKA attenuated tyrosine phosphorylation of the EGFR by angiotensin II stimulation, but this did not occur following direct stimulation of the EGFR with EGF, indicating that these proteins function between the activated AT1R and the EGFR. Further investigation of TRIO and CHKA revealed that their activity is likely to be required for AT1R-EGFR transactivation. CHKA also mediated EGFR transactivation in response to another G protein-coupled receptor (GPCR) ligand, thrombin, indicating a pervasive role for CHKA in GPCR-EGFR crosstalk. Our study reveals the power of unbiased, functional genomic screens to identify new signalling mediators important for tissue remodelling in cardiovascular disease and cancer.
authors.affiliation: School of Biomedical Sciences, The University of Queensland, St. Lucia, Queensland, 4072, Australia.
authors.forename: Amee J
authors.initials: AJ
authors.lastname: George
authors.forename: Brooke W
authors.initials: BW
authors.lastname: Purdue
authors.forename: Cathryn M
authors.initials: CM
authors.lastname: Gould
authors.forename: Daniel W
authors.initials: DW
authors.lastname: Thomas
authors.forename: Yanny
authors.initials: Y
authors.lastname: Handoko
authors.forename: Hongwei
authors.initials: H
authors.lastname: Qian
authors.forename: Gregory A
authors.initials: GA
authors.lastname: Quaife-Ryan
authors.forename: Kylie A
authors.initials: KA
authors.lastname: Morgan
authors.forename: Kaylene J
authors.initials: KJ
authors.lastname: Simpson
authors.forename: Walter G
authors.initials: WG
authors.lastname: Thomas
authors.forename: Ross D
authors.initials: RD
authors.lastname: Hannan
mesh_terms: D000804:Angiotensin II / Q000378:metabolism / Q000494:pharmacology; D002461:Cell Line, Transformed; D002797:Choline Kinase / Q000037:antagonists & inhibitors / Q000235:genetics / Q000378:metabolism; D004815:Epidermal Growth Factor / Q000378:metabolism / Q000494:pharmacology; D004847:Epithelial Cells / Q000166:cytology / Q000187:drug effects / Q000378:metabolism*; D066246:ErbB Receptors / Q000235:genetics / Q000378:metabolism*; D005260:Female; D020662:Guanine Nucleotide Exchange Factors / Q000037:antagonists & inhibitors / Q000235:genetics / Q000378:metabolism; D006801:Humans; D042361:Mammary Glands, Human / Q000166:cytology / Q000187:drug effects / Q000378:metabolism*; D017346:Protein Serine-Threonine Kinases / Q000037:antagonists & inhibitors / Q000235:genetics / Q000378:metabolism; D011505:Protein-Tyrosine Kinases / Q000037:antagonists & inhibitors / Q000235:genetics / Q000378:metabolism; D034741:RNA, Small Interfering / Q000235:genetics / Q000378:metabolism; D044140:Receptor, Angiotensin, Type 1 / Q000235:genetics / Q000378:metabolism*; D015398:Signal Transduction; D013917:Thrombin / Q000378:metabolism / Q000494:pharmacology; D015533:Transcriptional Activation
chemical_list: D020662:Guanine Nucleotide Exchange Factors; D034741:RNA, Small Interfering; D044140:Receptor, Angiotensin, Type 1; D000804:Angiotensin II; D004815:Epidermal Growth Factor; C492367:BMX protein, human; C517212:CHKA protein, human; D002797:Choline Kinase; C512478:EGFR protein, human; D066246:ErbB Receptors; D011505:Protein-Tyrosine Kinases; D017346:Protein Serine-Threonine Kinases; C099628:TRIO protein, human; D013917:Thrombin
keywords: Angiotensin; EGFR; G protein-coupled receptor; siRNA, T</emph>ransactivation
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