Herring B P, Smith A F
Department of Physiology and Biophysics, Indiana University School of Medicine, Indianapolis 46202-5120, USA.
Am J Physiol. 1996 Jun;270(6 Pt 1):C1656-65. doi: 10.1152/ajpcell.1996.270.6.C1656.
The carboxy terminus of the smooth muscle myosin light chain kinase (smMLCK) is expressed as an independent protein, telokin. Western and Northern blotting analyses demonstrated that telokin protein and mRNA are expressed at high levels only in adult and embryonic smooth muscle tissues and cells. In vitro transfection assays in A10 smooth muscle cells identified a functional promoter located in an intron in the 3' region of the smMLCK gene that directs the smooth muscle cell-specific transcription of telokin. To test the cell specificity of the telokin promoter in vivo, transgenic mice were generated in which the telokin promoter was used to drive expression of SV40 large T-antigen. Expression of T-antigen in the transgenic mice paralleled that of the endogenous telokin gene. High levels of T-antigen expression were observed in smooth muscle tissues of the digestive, urinary, and reproductive tracts, with lower levels of expression in airway and vascular smooth muscle. Expression was restricted to smooth muscle cells, with no expression detected in any other cell type.
平滑肌肌球蛋白轻链激酶(smMLCK)的羧基末端作为一种独立的蛋白——端激酶表达。蛋白质免疫印迹和Northern印迹分析表明,端激酶蛋白和mRNA仅在成年和胚胎平滑肌组织及细胞中高水平表达。在A10平滑肌细胞中进行的体外转染试验确定了一个位于smMLCK基因3'区域内含子中的功能性启动子,该启动子指导端激酶的平滑肌细胞特异性转录。为了在体内测试端激酶启动子的细胞特异性,构建了转基因小鼠,其中端激酶启动子用于驱动SV40大T抗原的表达。转基因小鼠中T抗原的表达与内源性端激酶基因的表达相似。在消化、泌尿和生殖道的平滑肌组织中观察到高水平的T抗原表达,在气道和血管平滑肌中的表达水平较低。表达仅限于平滑肌细胞,在任何其他细胞类型中均未检测到表达。