title: Dynamic and coordinated single-molecular interactions at TM4SF5-enriched microdomains guide invasive behaviors in 2- and 3-dimensional environments.
abstract: Membrane proteins sense extracellular cues and transduce intracellular signaling to coordinate directionality and speed during cellular migration. They are often localized to specific regions, as with lipid rafts or tetraspanin-enriched microdomains; however, the dynamic interactions of tetraspanins with diverse receptors within tetraspanin-enriched microdomains on cellular surfaces remain largely unexplored. Here, we investigated effects of tetraspan(in) TM4SF5 (transmembrane 4 L6 family member 5)-enriched microdomains (T5ERMs) on the directionality of cell migration. Physical association of TM4SF5 with epidermal growth factor receptor (EGFR) and integrin α5 was visualized by live fluorescence cross-correlation spectroscopy and higher-resolution microscopy at the leading edge of migratory cells, presumably forming TM4SF5-enriched microdomains. Whereas TM4SF5 and EGFR colocalized at the migrating leading region more than at the rear, TM4SF5 and integrin α5 colocalized evenly throughout cells. Cholesterol depletion and disruption in TM4SF5 post-translational modifications, includingN-glycosylation and palmitoylation, altered TM4SF5 interactions and cellular localization, which led to less cellular migration speed and directionality in 2- or 3-dimensional conditions. TM4SF5 controlled directional cell migration and invasion, and importantly, these TM4SF5 functions were dependent on cholesterol, TM4SF5 post-translational modifications, and EGFR and integrin α5 activity. Altogether, we showed that TM4SF5 dynamically interacted with EGFR and integrin α5 in migratory cells to control directionality and invasion.-Kim, H.-J., Kwon, S., Nam, S. H., Jung, J. W., Kang, M., Ryu, J., Kim, J. E., Cheong, J.-G., Cho, C. Y., Kim, S., Song, D.-G., Kim, Y.-N., Kim, T. Y., Jung, M.-K., Lee, K.-M., Pack, C.-G., Lee, J. W. Dynamic and coordinated single-molecular interactions at TM4SF5-enriched microdomains guide invasive behaviors in 2- and 3-dimensional environments.
authors.affiliation: Department of Pharmacy, Research Institute of Pharmaceutical Sciences, College of Pharmacy, Seoul National University, Seoul, South Korea.
authors.forename: Hye-Jin
authors.initials: HJ
authors.lastname: Kim
authors.affiliation: Department of Pharmacy, Research Institute of Pharmaceutical Sciences, College of Pharmacy, Seoul National University, Seoul, South Korea.
authors.forename: Sojung
authors.initials: S
authors.lastname: Kwon
authors.affiliation: Interdisciplinary Program in Genetic Engineering, Seoul National University, Seoul, South Korea.
authors.forename: Seo Hee
authors.initials: SH
authors.lastname: Nam
authors.affiliation: Interdisciplinary Program in Genetic Engineering, Seoul National University, Seoul, South Korea.
authors.forename: Jae Woo
authors.initials: JW
authors.lastname: Jung
authors.affiliation: Department of Pharmacy, Research Institute of Pharmaceutical Sciences, College of Pharmacy, Seoul National University, Seoul, South Korea.
authors.forename: Minkyung
authors.initials: M
authors.lastname: Kang
authors.affiliation: Department of Pharmacy, Research Institute of Pharmaceutical Sciences, College of Pharmacy, Seoul National University, Seoul, South Korea.
authors.forename: Jihye
authors.initials: J
authors.lastname: Ryu
authors.affiliation: Department of Pharmacy, Research Institute of Pharmaceutical Sciences, College of Pharmacy, Seoul National University, Seoul, South Korea.
authors.forename: Ji Eon
authors.initials: JE
authors.lastname: Kim
authors.affiliation: Department of Pharmacy, Research Institute of Pharmaceutical Sciences, College of Pharmacy, Seoul National University, Seoul, South Korea.
authors.forename: Jin-Gyu
authors.initials: JG
authors.lastname: Cheong
authors.affiliation: Department of Pharmacy, Research Institute of Pharmaceutical Sciences, College of Pharmacy, Seoul National University, Seoul, South Korea.
authors.forename: Chang Yun
authors.initials: CY
authors.lastname: Cho
authors.affiliation: Department of Pharmacy, Research Institute of Pharmaceutical Sciences, College of Pharmacy, Seoul National University, Seoul, South Korea.
authors.forename: Somi
authors.initials: S
authors.lastname: Kim
authors.affiliation: Department of Pharmacy, Research Institute of Pharmaceutical Sciences, College of Pharmacy, Seoul National University, Seoul, South Korea.
authors.forename: Dae-Geun
authors.initials: DG
authors.lastname: Song
authors.affiliation: National Cancer Center, Gyeonggi-do, South Korea.
authors.forename: Yong-Nyun
authors.initials: YN
authors.lastname: Kim
authors.affiliation: Department of Pharmacy, Research Institute of Pharmaceutical Sciences, College of Pharmacy, Seoul National University, Seoul, South Korea.
authors.forename: Tai Young
authors.initials: TY
authors.lastname: Kim
authors.affiliation: Department of Convergence Medicine, University of Ulsan College of Medicine and Asan Institute for Life Sciences, Asan Medical Center, Seoul, South Korea.
authors.forename: Min-Kyo
authors.initials: MK
authors.lastname: Jung
authors.affiliation: Department of Pharmacy, Research Institute of Pharmaceutical Sciences, College of Pharmacy, Seoul National University, Seoul, South Korea.
authors.forename: Kyung-Min
authors.initials: KM
authors.lastname: Lee
authors.affiliation: Department of Convergence Medicine, University of Ulsan College of Medicine and Asan Institute for Life Sciences, Asan Medical Center, Seoul, South Korea.
authors.forename: Chan-Gi
authors.initials: CG
authors.lastname: Pack
authors.affiliation: Department of Pharmacy, Research Institute of Pharmaceutical Sciences, College of Pharmacy, Seoul National University, Seoul, South Korea; jwl@snu.ac.kr.|Interdisciplinary Program in Genetic Engineering, Seoul National University, Seoul, South Korea.
authors.forename: Jung Weon
authors.initials: JW
authors.lastname: Lee
mesh_terms: D045744:Cell Line, Tumor; D002465:Cell Movement; D002784:Cholesterol / Q000378:metabolism; D066246:ErbB Receptors / Q000378:metabolism; D006031:Glycosylation; D057809:HEK293 Cells; D022781:Hepatocytes / Q000378:metabolism / Q000502:physiology / Q000648:ultrastructure; D006801:Humans; D039482:Integrin alpha5 / Q000378:metabolism; D054878:Lipoylation; D021962:Membrane Microdomains / Q000378:metabolism* / Q000648:ultrastructure; D008565:Membrane Proteins / Q000378:metabolism*; D011485:Protein Binding; D011499:Protein Processing, Post-Translational
chemical_list: D039482:Integrin alpha5; D008565:Membrane Proteins; C111029:TM4SF5 protein, human; D002784:Cholesterol; D066246:ErbB Receptors
keywords: 3D cell culture; FCCS; membrane proteins; migration; protein interaction affinity