title: EGF-stimulated activation of Rab35 regulates RUSC2-GIT2 complex formation to stabilize GIT2 during directional lung cancer cell migration.
abstract: Non-small cell lung cancer (NSCLC) remains one of the most metastasizing tumors, and directional cell migration is critical for targeting tumor metastasis. GIT2 has been known to bind to Paxillin to control cell polarization and directional migration. However, the molecular mechanisms underlying roles of GIT2 in controlling cell polarization and directional migration remain elusive. Here we demonstrated GIT2 control cell polarization and direction dependent on the regulation of Golgi through RUSC2. RUSC2 interacts with SHD of GIT2 in various lung cancer cells, and stabilizes GIT2 (Mazaki et al., 2006; Yu et al., 2009) by decreasing degradation and increasing its phosphorylation. Silencing of RUSC2 showed reduced stability of GIT2, defective Golgi reorientation toward the wound edge and decreased directional migration. Moreover, short-term EGF stimulation can increase the interaction between RUSC2 and GIT2, prolonged stimulation leads to a decrease of their interaction through activating Rab35. Silencing of Rab35 also reduced stability and phosphorylation of GIT2 and decreased cell migration. Taken together, our study indicated that RUSC2 participates in EGFR signaling and regulates lung cancer progression, and may be a new therapeutic target against lung cancer metastasis.
authors.affiliation: Department of Physiology, Nanjing Medical University, Nanjing, Jiangsu 210029, China.
authors.forename: Biao
authors.initials: B
authors.lastname: Duan
authors.affiliation: Department of Biochemistry and Molecular Biology, Nanjing Medical University, Nanjing, Jiangsu 210029, China; Department of Physiology, Xuzhou Medical University, Xuzhou, Jiangsu 221004, China.
authors.forename: Jie
authors.initials: J
authors.lastname: Cui
authors.affiliation: Department of Physiology, Nanjing Medical University, Nanjing, Jiangsu 210029, China.
authors.forename: Shixiu
authors.initials: S
authors.lastname: Sun
authors.affiliation: Department of Physiology, Nanjing Medical University, Nanjing, Jiangsu 210029, China.
authors.forename: Jianchao
authors.initials: J
authors.lastname: Zheng
authors.affiliation: Department of Biochemistry and Molecular Biology, Nanjing Medical University, Nanjing, Jiangsu 210029, China; Jiangsu Key Lab of Cancer Biomarkers, Prevention and Treatment, Collaborative Innovation Center For Cancer Personalized Medicine, Nanjing Medical University, Nanjing, Jiangsu 211166, China.
authors.forename: Yujie
authors.initials: Y
authors.lastname: Zhang
authors.affiliation: Department of Gastroenterology, The First Affiliated Hospital of Nanjing Medical University, Nanjing, Jiangsu 210029, China.
authors.forename: Bixing
authors.initials: B
authors.lastname: Ye
authors.affiliation: Department of Biochemistry and Molecular Biology, Nanjing Medical University, Nanjing, Jiangsu 210029, China.
authors.forename: Yan
authors.initials: Y
authors.lastname: Chen
authors.affiliation: Department of Physiology, Nanjing Medical University, Nanjing, Jiangsu 210029, China.
authors.forename: Wenjie
authors.initials: W
authors.lastname: Deng
authors.affiliation: Department of Physiology, Nanjing Medical University, Nanjing, Jiangsu 210029, China; Jiangsu Key Lab of Cancer Biomarkers, Prevention and Treatment, Collaborative Innovation Center For Cancer Personalized Medicine, Nanjing Medical University, Nanjing, Jiangsu 211166, China.
authors.forename: Jun
authors.initials: J
authors.lastname: Du
authors.affiliation: Department of Physiology, Nanjing Medical University, Nanjing, Jiangsu 210029, China.
authors.forename: Yichao
authors.initials: Y
authors.lastname: Zhu
authors.affiliation: Department of Physiology, School of Medicine, Jiangsu University, Zhenjiang, Jiangsu 212013, China.
authors.forename: Yongchang
authors.initials: Y
authors.lastname: Chen
authors.affiliation: Department of Physiology, Nanjing Medical University, Nanjing, Jiangsu 210029, China; Department of Biochemistry and Molecular Biology, Nanjing Medical University, Nanjing, Jiangsu 210029, China; Jiangsu Key Lab of Cancer Biomarkers, Prevention and Treatment, Collaborative Innovation Center For Cancer Personalized Medicine, Nanjing Medical University, Nanjing, Jiangsu 211166, China. Electronic address: lgu@njmu.edu.cn.
authors.forename: Luo
authors.initials: L
authors.lastname: Gu
mesh_terms: D000072283:A549 Cells; D002289:Carcinoma, Non-Small-Cell Lung / Q000201:enzymology* / Q000235:genetics / Q000473:pathology; D002352:Carrier Proteins / Q000235:genetics / Q000378:metabolism*; D002465:Cell Movement / Q000187:drug effects*; D004789:Enzyme Activation; D004815:Epidermal Growth Factor / Q000494:pharmacology*; D066246:ErbB Receptors / Q000819:agonists / Q000378:metabolism; D020690:GTPase-Activating Proteins / Q000235:genetics / Q000378:metabolism*; D006056:Golgi Apparatus / Q000187:drug effects / Q000378:metabolism; D057809:HEK293 Cells; D006801:Humans; D008175:Lung Neoplasms / Q000201:enzymology* / Q000235:genetics / Q000473:pathology; D010766:Phosphorylation; D011485:Protein Binding; D054730:Protein Interaction Domains and Motifs; D055550:Protein Stability; D021381:Protein Transport; D034622:RNA Interference; D015398:Signal Transduction / Q000187:drug effects; D013997:Time Factors; D014162:Transfection; D020691:rab GTP-Binding Proteins / Q000235:genetics / Q000378:metabolism*; D018909:src Homology Domains
chemical_list: D002352:Carrier Proteins; C430213:GIT2 protein, human; D020690:GTPase-Activating Proteins; C509346:RUSC2 protein, human; D004815:Epidermal Growth Factor; C512478:EGFR protein, human; D066246:ErbB Receptors; C511183:RAB35 protein, human; D020691:rab GTP-Binding Proteins
keywords: Directional migration; GIT2; Non-small cell lung cancer; Protein degradation; RUSC2