title: Association between α4 integrin cytoplasmic tail and non-muscle myosin IIA regulates cell migration.
abstract: α4β1 integrin regulates cell migration via cytoplasmic interactions. Here, we report an association between the cytoplasmic tail of α4 integrin (α4 tail) and non-muscle myosin IIA (MIIA), demonstrated by co-immunoprecipitation of the MIIA heavy chain (HC) with anti-α4-integrin antibodies and pull-down of MIIA-HC with recombinant α4 tail from cell lysates. The association between the α4 tail and MIIA does not require paxillin binding or phosphorylation at Ser988 in the α4 tail. We found that substituting Glu982 in the α4 tail with alanine (E982A) disrupts the α4-MIIA association without interfering with the paxillin binding or Ser988 phosphorylation. By comparing stably transfected CHO cells, we show that the E982A mutation reduces the ability of α4β1 integrin to mediate cell spreading and to promote front-back polarization. In addition, we show that E982A impairs shear-flow-induced migration of the α4-integrin-expressing CHO cells by reducing their migration speed and directional persistence. The E982A mutation also leads to defects in the organization of MIIA filament bundles. Furthermore, when cells are plated on fibronectin and simulated with shear flow, α4β1 integrin forms filament-like patterns that co-align with MIIA filament bundles. These results provide a new mechanism for linking integrins to the actomyosin cytoskeleton and for regulating cell migration by integrins and non-muscle myosin II.
authors.affiliation: Department of Cell Biology, The Johns Hopkins University School of Medicine, Baltimore, MD 21205, USA.
authors.forename: Leslie A
authors.initials: LA
authors.lastname: Rivera Rosado
authors.forename: Troy A
authors.initials: TA
authors.lastname: Horn
authors.forename: Sara C
authors.initials: SC
authors.lastname: McGrath
authors.forename: Robert J
authors.initials: RJ
authors.lastname: Cotter
authors.forename: Joy T
authors.initials: JT
authors.lastname: Yang
mesh_terms: D019943:Amino Acid Substitution; D000818:Animals; D016466:CHO Cells; D002465:Cell Movement / Q000502:physiology*; D006224:Cricetinae; D003412:Cricetulus; D006801:Humans; D039441:Integrin alpha4 / Q000378:metabolism*; D039041:Integrin alpha4beta1 / Q000378:metabolism*; D051379:Mice; D024507:Nonmuscle Myosin Type IIA / Q000378:metabolism*; D051419:Paxillin / Q000378:metabolism; D011485:Protein Binding
chemical_list: D039041:Integrin alpha4beta1; D051419:Paxillin; D039441:Integrin alpha4; D024507:Nonmuscle Myosin Type IIA