Tissue-selective expression of a conditionally-active ROCK2-estrogen receptor fusion protein

Michael S. Samuel, Nicola Rath, Siti Fathiah Masre, Sarah T. Boyle, David A. Greenhalgh, Marina Kochetkova, Sheila Bryson, David Stevenson, Michael F. Olson

Research output: Contribution to journalArticle

2 Citations (Scopus)

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.

Original languageEnglish
Pages (from-to)636-646
Number of pages11
JournalGenesis
Volume54
Issue number12
DOIs
Publication statusPublished - 1 Dec 2016

Fingerprint

Phosphotransferases
rhoA GTP-Binding Protein
Gammaretrovirus
Estradiol Congeners
Actomyosin
Recombinases
Protein-Serine-Threonine Kinases
Tamoxifen
Intestinal Mucosa
Research
Epidermis
Estrogen Receptors
Cytochrome P-450 Enzyme System
Breast
Cell Proliferation
Hormones
Breast Neoplasms
Hypertension
Genes
estrophilin

Keywords

  • actomyosin contractility
  • Hprt locus
  • ROCK

ASJC Scopus subject areas

  • Genetics
  • Endocrinology
  • Cell Biology

Cite this

Samuel, M. S., Rath, N., Masre, S. F., Boyle, S. T., Greenhalgh, D. A., Kochetkova, M., ... Olson, M. F. (2016). Tissue-selective expression of a conditionally-active ROCK2-estrogen receptor fusion protein. Genesis, 54(12), 636-646. https://doi.org/10.1002/dvg.22988

Tissue-selective expression of a conditionally-active ROCK2-estrogen receptor fusion protein. / Samuel, Michael S.; Rath, Nicola; Masre, Siti Fathiah; Boyle, Sarah T.; Greenhalgh, David A.; Kochetkova, Marina; Bryson, Sheila; Stevenson, David; Olson, Michael F.

In: Genesis, Vol. 54, No. 12, 01.12.2016, p. 636-646.

Research output: Contribution to journalArticle

Samuel, MS, Rath, N, Masre, SF, Boyle, ST, Greenhalgh, DA, Kochetkova, M, Bryson, S, Stevenson, D & Olson, MF 2016, 'Tissue-selective expression of a conditionally-active ROCK2-estrogen receptor fusion protein', Genesis, vol. 54, no. 12, pp. 636-646. https://doi.org/10.1002/dvg.22988
Samuel MS, Rath N, Masre SF, Boyle ST, Greenhalgh DA, Kochetkova M et al. Tissue-selective expression of a conditionally-active ROCK2-estrogen receptor fusion protein. Genesis. 2016 Dec 1;54(12):636-646. https://doi.org/10.1002/dvg.22988
Samuel, Michael S. ; Rath, Nicola ; Masre, Siti Fathiah ; Boyle, Sarah T. ; Greenhalgh, David A. ; Kochetkova, Marina ; Bryson, Sheila ; Stevenson, David ; Olson, Michael F. / Tissue-selective expression of a conditionally-active ROCK2-estrogen receptor fusion protein. In: Genesis. 2016 ; Vol. 54, No. 12. pp. 636-646.
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