title: Tissue-selective expression of a conditionally-active ROCK2-estrogen receptor fusion protein.
abstract: The serine/threonine kinases ROCK1 and ROCK2 are central mediators of actomyosin contractile force generation that act downstream of the RhoA small GTP-binding protein. As a result, they have key roles in regulating cell morphology and proliferation, and have been implicated in numerous pathological conditions and diseases including hypertension and cancer. Here we describe the generation of a gene-targeted mouse line that enables CRE-inducible expression of a conditionally-active fusion between the ROCK2 kinase domain and the hormone-binding domain of a mutated estrogen receptor (ROCK2:ER). This two-stage system of regulation allows for tissue-selective expression of the ROCK2:ER fusion protein, which then requires administration of estrogen analogues such as tamoxifen or 4-hydroxytamoxifen to elicit kinase activity. This conditional gain-of-function system was validated in multiple tissues by crossing with mice expressing CRE recombinase under the transcriptional control of cytokeratin14 (K14), murine mammary tumor virus (MMTV) or cytochrome P450 Cyp1A1 (Ah) promoters, driving appropriate expression in the epidermis, mammary or intestinal epithelia respectively. Given the interest in ROCK signaling in normal physiology and disease, this mouse line will facilitate research into the consequences of ROCK activation that could be used to complement conditional knockout models. Birth Defects Research (Part A) 106:636-646, 2016. © 2016 Wiley Periodicals, Inc.
authors: Centre for Cancer Biology, SA Pathology and the University of South Australia, Frome Road, Adelaide, South Australia, 5000, Australia.|School of Medicine, Faculty of Health Sciences, University of Adelaide, Adelaide, South Australia, 5000, Australia.
authors: Michael S
authors: MS
authors: Samuel
authors: Cancer Research UK Beatson Institute, Garscube Estate, Switchback Road, Glasgow, G61 1BD, UK.
authors: Nicola
authors: N
authors: Rath
authors: Biomedical Science Programme, School of Diagnostic and Applied Health Sciences, Faculty of Allied Health Sciences, University of Kebangsaan, Kuala Lumpur, 50300, Malaysia.|Section of Dermatology and Molecular Carcinogenesis College of Medical Veterinary and Life Sciences, University of Glasgow, Glasgow, G12 8QQ, UK.
authors: Siti F
authors: SF
authors: Masre
authors: Centre for Cancer Biology, SA Pathology and the University of South Australia, Frome Road, Adelaide, South Australia, 5000, Australia.
authors: Sarah T
authors: ST
authors: Boyle
authors: Section of Dermatology and Molecular Carcinogenesis College of Medical Veterinary and Life Sciences, University of Glasgow, Glasgow, G12 8QQ, UK.
authors: David A
authors: DA
authors: Greenhalgh
authors: Centre for Cancer Biology, SA Pathology and the University of South Australia, Frome Road, Adelaide, South Australia, 5000, Australia.
authors: Marina
authors: M
authors: Kochetkova
authors: Cancer Research UK Beatson Institute, Garscube Estate, Switchback Road, Glasgow, G61 1BD, UK.
authors: Sheila
authors: S
authors: Bryson
authors: Cancer Research UK Beatson Institute, Garscube Estate, Switchback Road, Glasgow, G61 1BD, UK.
authors: David
authors: D
authors: Stevenson
authors: Cancer Research UK Beatson Institute, Garscube Estate, Switchback Road, Glasgow, G61 1BD, UK.|Institute of Cancer Sciences, University of Glasgow, Glasgow, G12 8QQ, UK.
authors: Michael F
authors: MF
authors: Olson
mesh_terms: D000818:Animals; D019363:Cytochrome P-450 CYP1A1 / Q000235:genetics; D004817:Epidermis / Q000378:metabolism; D047628:Estrogen Receptor alpha / Q000096:biosynthesis / Q000235:genetics*; D005260:Female; D018507:Gene Expression Regulation, Developmental / Q000187:drug effects; D006801:Humans; D019426:Integrases / Q000235:genetics; D007413:Intestinal Mucosa / Q000378:metabolism; D008321:Mammary Glands, Animal / Q000378:metabolism; D051379:Mice; D009928:Organ Specificity / Q000235:genetics; D011401:Promoter Regions, Genetic / Q000235:genetics; D011993:Recombinant Fusion Proteins / Q000096:biosynthesis / Q000235:genetics*; D015398:Signal Transduction / Q000187:drug effects; D013629:Tamoxifen / Q000008:administration & dosage; D054460:rho-Associated Kinases / Q000096:biosynthesis / Q000235:genetics*
chemical_list: D047628:Estrogen Receptor alpha; D011993:Recombinant Fusion Proteins; D013629:Tamoxifen; D019363:Cytochrome P-450 CYP1A1; C515855:Rock2 protein, mouse; D054460:rho-Associated Kinases; C045073:Cre recombinase; D019426:Integrases
keywords: Hprt locus; ROCK; actomyosin contractility