title: PKA and actin play critical roles as downstream effectors in MRP4-mediated regulation of fibroblast migration.
abstract: Multidrug resistance protein 4 (MRP4), a member of the ATP binding cassette transporter family, functions as a plasma membrane exporter of cyclic nucleotides. Recently, we demonstrated that fibroblasts lacking the Mrp4 gene migrate faster and contain higher cyclic-nucleotide levels. Here, we show that cAMP accumulation and protein kinase A (PKA) activity are higher and polarized in Mrp4(-/-) fibroblasts, versus Mrp4(+/+) cells. MRP4-containing macromolecular complexes isolated from these fibroblasts contained several proteins, including actin, which play important roles in cell migration. We found that actin interacts with MRP4, predominantly at the plasma membrane, and an intact actin cytoskeleton is required to restrict MRP4 to specific microdomains of the plasma membrane. Our data further indicated that the enhanced accumulation of cAMP in Mrp4(-/-) fibroblasts facilitates cortical actin polymerization in a PKA-dependent manner at the leading edge, which in turn increases the overall rate of cell migration to accelerate the process of wound healing. Disruption of actin polymerization or inhibition of PKA activity abolished the effect of MRP4 on cell migration. Together, our findings suggest a novel cAMP-dependent mechanism for MRP4-mediated regulation of fibroblast migration whereby PKA and actin play critical roles as downstream effectors.
authors.affiliation: Division of Pulmonary Medicine, Department of Pediatrics, Cincinnati Children's Hospital Medical Center, Cincinnati, OH 45229, USA; Department of Physiology, University of Tennessee Health Science Center, Memphis, TN 38163, USA.
authors.forename: Chandrima
authors.initials: C
authors.lastname: Sinha
authors.affiliation: Department of Physiology, University of Tennessee Health Science Center, Memphis, TN 38163, USA.
authors.forename: Aixia
authors.initials: A
authors.lastname: Ren
authors.affiliation: Division of Pulmonary Medicine, Department of Pediatrics, Cincinnati Children's Hospital Medical Center, Cincinnati, OH 45229, USA.
authors.forename: Kavisha
authors.initials: K
authors.lastname: Arora
authors.affiliation: Division of Pulmonary Medicine, Department of Pediatrics, Cincinnati Children's Hospital Medical Center, Cincinnati, OH 45229, USA.
authors.forename: Chang Suk
authors.initials: CS
authors.lastname: Moon
authors.affiliation: Division of Pulmonary Medicine, Department of Pediatrics, Cincinnati Children's Hospital Medical Center, Cincinnati, OH 45229, USA.
authors.forename: Sunitha
authors.initials: S
authors.lastname: Yarlagadda
authors.affiliation: Division of Pulmonary Medicine, Department of Pediatrics, Cincinnati Children's Hospital Medical Center, Cincinnati, OH 45229, USA.
authors.forename: Koryse
authors.initials: K
authors.lastname: Woodrooffe
authors.affiliation: Department of Pediatrics, Emory University, Atlanta, GA 30322, USA.
authors.forename: Songbai
authors.initials: S
authors.lastname: Lin
authors.affiliation: Department of Pharmaceutical Sciences, St. Jude Children's Research Hospital, Memphis, TN, USA.
authors.forename: John D
authors.initials: JD
authors.lastname: Schuetz
authors.affiliation: Department of Pediatrics, Emory University, Atlanta, GA 30322, USA.
authors.forename: Assem G
authors.initials: AG
authors.lastname: Ziady
authors.affiliation: Division of Pulmonary Medicine, Department of Pediatrics, Cincinnati Children's Hospital Medical Center, Cincinnati, OH 45229, USA; Department of Physiology, University of Tennessee Health Science Center, Memphis, TN 38163, USA. Electronic address: anaren@cchmc.org.
authors.forename: Anjaparavanda P
authors.initials: AP
authors.lastname: Naren
mesh_terms: D008841:Actin Cytoskeleton / Q000187:drug effects / Q000378:metabolism; D000199:Actins / Q000737:chemistry / Q000378:metabolism*; D000818:Animals; D002460:Cell Line; D002462:Cell Membrane / Q000378:metabolism; D002465:Cell Movement / Q000187:drug effects; D000242:Cyclic AMP / Q000378:metabolism; D017868:Cyclic AMP-Dependent Protein Kinases / Q000378:metabolism*; D005347:Fibroblasts / Q000166:cytology / Q000187:drug effects / Q000378:metabolism; D057809:HEK293 Cells; D006801:Humans; D047468:Immunoprecipitation; D051379:Mice; D008856:Microscopy, Fluorescence; D027425:ATP-Binding Cassette, Sub-Family C Proteins / Q000037:antagonists & inhibitors / Q000235:genetics / Q000378:metabolism*; D041681:NIH 3T3 Cells; D011422:Propionates / Q000633:toxicity; D011485:Protein Binding; D011804:Quinolines / Q000633:toxicity
chemical_list: D000199:Actins; D000242:Cyclic AMP; D017868:Cyclic AMP-Dependent Protein Kinases; D027425:ATP-Binding Cassette, Sub-Family C Proteins; D011422:Propionates; D011804:Quinolines; C073492:ABCC4 protein, human; C059141:verlukast
keywords: Actin; Cyclic nucleotides; MRP4; Migration; PKA
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