title: Knockout of the PKN Family of Rho Effector Kinases Reveals a Non-redundant Role for PKN2 in Developmental Mesoderm Expansion.
abstract: In animals, the protein kinase C (PKC) family has expanded into diversely regulated subgroups, including the Rho family-responsive PKN kinases. Here, we describe knockouts of all three mouse PKN isoforms and reveal that PKN2 loss results in lethality at embryonic day 10 (E10), with associated cardiovascular and morphogenetic defects. The cardiovascular phenotype was not recapitulated by conditional deletion of PKN2 in endothelial cells or the developing heart. In contrast, inducible systemic deletion of PKN2 after E7 provoked collapse of the embryonic mesoderm. Furthermore, mouse embryonic fibroblasts, which arise from the embryonic mesoderm, depend on PKN2 for proliferation and motility. These cellular defects are reflected in vivo as dependence on PKN2 for mesoderm proliferation and neural crest migration. We conclude that failure of the mesoderm to expand in the absence of PKN2 compromises cardiovascular integrity and development, resulting in lethality.
authors.affiliation: Kinase Biology Laboratory, John Vane Science Centre, Barts Cancer Institute, Queen Mary University of London, Charterhouse Square, London EC1M 6BQ, UK.
authors.forename: Ivan
authors.initials: I
authors.lastname: Quétier
authors.affiliation: Protein Phosphorylation Laboratory, Francis Crick Institute, 44 Lincoln's Inn Fields, London WC2A 3LY, UK.
authors.forename: Jacqueline J T
authors.initials: JJT
authors.lastname: Marshall
authors.affiliation: Francis Crick Institute, 44 Lincoln's Inn Fields, London WC2A 3LY, UK.
authors.forename: Bradley
authors.initials: B
authors.lastname: Spencer-Dene
authors.affiliation: Protein Phosphorylation Laboratory, Francis Crick Institute, 44 Lincoln's Inn Fields, London WC2A 3LY, UK.
authors.forename: Sylvie
authors.initials: S
authors.lastname: Lachmann
authors.affiliation: Protein Phosphorylation Laboratory, Francis Crick Institute, 44 Lincoln's Inn Fields, London WC2A 3LY, UK.
authors.forename: Adele
authors.initials: A
authors.lastname: Casamassima
authors.affiliation: Instituto Medicina Molecular (iMM), Faculdade de Medicina da Universidade de Lisboa, 1649-028 Lisbon, Portugal.
authors.forename: Claudio
authors.initials: C
authors.lastname: Franco
authors.affiliation: Newlife Birth Defects Research Centre, Institute of Child Health, University College, London WC1N 1EH, UK.
authors.forename: Sarah
authors.initials: S
authors.lastname: Escuin
authors.affiliation: John Vane Science Centre, Barts Cancer Institute, Queen Mary University of London, Charterhouse Square, London EC1M 6BQ, UK.
authors.forename: Joseph T
authors.initials: JT
authors.lastname: Worrall
authors.affiliation: Kinase Biology Laboratory, John Vane Science Centre, Barts Cancer Institute, Queen Mary University of London, Charterhouse Square, London EC1M 6BQ, UK.
authors.forename: Priththivika
authors.initials: P
authors.lastname: Baskaran
authors.affiliation: John Vane Science Centre, Barts Cancer Institute, Queen Mary University of London, Charterhouse Square, London EC1M 6BQ, UK.
authors.forename: Vinothini
authors.initials: V
authors.lastname: Rajeeve
authors.affiliation: Francis Crick Institute, 44 Lincoln's Inn Fields, London WC2A 3LY, UK.
authors.forename: Michael
authors.initials: M
authors.lastname: Howell
authors.affiliation: Newlife Birth Defects Research Centre, Institute of Child Health, University College, London WC1N 1EH, UK.
authors.forename: Andrew J
authors.initials: AJ
authors.lastname: Copp
authors.affiliation: Francis Crick Institute, 44 Lincoln's Inn Fields, London WC2A 3LY, UK.
authors.forename: Gordon
authors.initials: G
authors.lastname: Stamp
authors.affiliation: Genetic Manipulation Services, Francis Crick Institute, Clare Hall, Herts EN6 3LD, UK.
authors.forename: Ian
authors.initials: I
authors.lastname: Rosewell
authors.affiliation: John Vane Science Centre, Barts Cancer Institute, Queen Mary University of London, Charterhouse Square, London EC1M 6BQ, UK.
authors.forename: Pedro
authors.initials: P
authors.lastname: Cutillas
authors.affiliation: Max-Delbrück-Center for Molecular Medicine, Robert-Rössle-Strasse 10, 13125 Berlin, Germany.
authors.forename: Holger
authors.initials: H
authors.lastname: Gerhardt
authors.affiliation: Protein Phosphorylation Laboratory, Francis Crick Institute, 44 Lincoln's Inn Fields, London WC2A 3LY, UK; Division of Cancer Studies, King's College London, New Hunt's House, Saint Thomas Street, London SE1 1UL, UK. Electronic address: peter.parker@crick.ac.uk.
authors.forename: Peter J
authors.initials: PJ
authors.lastname: Parker
authors.affiliation: Kinase Biology Laboratory, John Vane Science Centre, Barts Cancer Institute, Queen Mary University of London, Charterhouse Square, London EC1M 6BQ, UK. Electronic address: a.cameron@qmul.ac.uk.
authors.forename: Angus J M
authors.initials: AJM
authors.lastname: Cameron
mesh_terms: D000818:Animals; D000970:Antineoplastic Agents / Q000494:pharmacology; D017209:Apoptosis / Q000187:drug effects; D002460:Cell Line; D002465:Cell Movement / Q000187:drug effects; D049109:Cell Proliferation / Q000187:drug effects; D004622:Embryo, Mammalian / Q000166:cytology / Q000378:metabolism; D047108:Embryonic Development / Q000187:drug effects; D017930:Genes, Reporter; D006321:Heart / Q000254:growth & development; D008648:Mesoderm / Q000166:cytology / Q000378:metabolism*; D051379:Mice; D008810:Mice, Inbred C57BL; D018345:Mice, Knockout; D008855:Microscopy, Electron, Scanning; D009206:Myocardium / Q000378:metabolism / Q000473:pathology; D011493:Protein Kinase C / Q000172:deficiency / Q000235:genetics* / Q000378:metabolism; D013629:Tamoxifen / Q000031:analogs & derivatives / Q000494:pharmacology
chemical_list: D000970:Antineoplastic Agents; D013629:Tamoxifen; C016601:afimoxifene; C061570:protein kinase N; D011493:Protein Kinase C
keywords: 4-OHT; 4-hydroxytamoxifen; ES cells; Embryonic stem cells; MEF; NCCs; PKC; PKN; Protein kinase N; mouse embryonic fibroblasts; neural crest cells; protein kinase C