FRNK, the autonomously expressed C-terminal region of focal adhesion kinase, is uniquely regulated in vascular smooth muscle: analysis of expression in transgenic mice.
FRNK, the autonomously expressed carboxyl-terminal region of focal adhesion kinase (FAK), is expressed in tissues that are rich in vascular smooth muscle cells (VSMCs). Here we report the generation of transgenic mice harboring the putative FRNK promoter fused to LacZ and examine the promoter activity in situ via expression of beta-galactosidase. The transgenic mice exhibited expression of beta-galactosidase predominantly in arterial VSMCs in large and small blood vessels of major organs. Upregulation of beta-galactosidase activity was observed in tunica media following carotid injury, indicating that the FRNK promoter is activated in VSMCs in response to injury. Robust expression of beta-galactosidase in blood vessels was also detected in the developing embryo. However, expression was also observed in the midline, the nose and skin epidermis, indicating distinct transcriptional regulation of the FRNK promoter in embryogenesis. To analyze FRNK expression in vitro, we identified a 116 bp sequence in the FRNK promoter that was sufficient to function as an enhancer when fused to the minimal actin promoter and expressed in cultured smooth muscle cells. Mutation of AP-1 and NF-E2 binding consensus sequences within this element abrogated enhancer activity, supporting the involvement of this promoter element in VSMC expression of FRNK.
Hayasaka
Haruko
H
Department of Microbiology, University of Virginia Health System, Charlottesville, Virginia 22908-0734, USA.
Simon
Kate
K
Hershey
E Daniel
ED
Masumoto
Koh-Hei
KH
Parsons
J Thomas
JT
D000818:Animals; D000831:Animals, Newborn; D001483:Base Sequence; D002460:Cell Line; D004622:Embryo, Mammalian / Q000196:embryology / Q000378:metabolism; D005786:Gene Expression Regulation / Q000235:genetics*; D018507:Gene Expression Regulation, Developmental / Q000235:genetics; D016678:Genome; D007763:Lac Operon / Q000235:genetics; D008297:Male; D051379:Mice; D008822:Mice, Transgenic; D008969:Molecular Sequence Data; D009131:Muscle, Smooth, Vascular / Q000201:enzymology*; D011401:Promoter Regions, Genetic / Q000235:genetics