Zajdel Robert W, Denz Christopher R, Lee Sung, Dube Syamalima, Ehler Elisabeth, Perriard Evylene, Perriard Jean-Claude, Dube Dipak K
Department of Cell and Developmental Biology, Syracuse, New York, USA.
J Cell Biochem. 2003 Jun 1;89(3):427-39. doi: 10.1002/jcb.10504.
Tropomyosins are present in various muscle (skeletal, cardiac, and smooth) and non-muscle cells with different isoforms characteristic of specific cell types. We describe here a novel smooth/striated chimeric isoform that was expressed in developing chick heart in addition to the classically described TM-4 type. This novel alpha-Tm tropomyosin isoform, designated as alpha-Tm-2, contains exon 2a (in place of exon 2b). The known striated muscle isoform (alpha-Tm-1) was also expressed in embryonic hearts along with the striated muscle isoform of TM-4. In adult heart, TM-4 was expressed, however, expression of both alpha-Tm-1 and alpha-Tm-2 isoforms was drastically reduced or downregulated. Interestingly, we were unable to detect the expression of alpha-Tm-2 in embryonic and adult skeletal muscle, however, the alpha-Tm-1 isoform is expressed in embryonic and adult skeletal muscle. Examination of other possible isoforms of the alpha-TM gene, i.e., alpha-smooth muscle tropomyosin (alpha-Sm), alpha-Fibroblast-1 (alpha-F1), and alpha-Fibroblast-2 (alpha-F2) revealed expression in embryonic hearts and a significant reduction of each of these isoforms in adult heart. In order to elucidate the role of the newly discovered tropomyosin isoform in chicken, we ectopically expressed the GFP fusion protein of alpha-Tm-1 and alpha-Tm-2 separately into cardiomyocytes isolated from neonatal rats. Each isoform was incorporated into organized myofibrils. Our results suggest that the alpha-TM gene may undergo both positive and negative transcriptional control in chicken hearts during development.
原肌球蛋白存在于各种肌肉(骨骼肌、心肌和平滑肌)以及非肌肉细胞中,具有特定细胞类型所特有的不同同工型。我们在此描述了一种新型的平滑肌/横纹肌嵌合同工型,除了经典描述的TM-4型外,它还在发育中的鸡心脏中表达。这种新型的α-Tm原肌球蛋白同工型,命名为α-Tm-2,包含外显子2a(代替外显子2b)。已知的横纹肌同工型(α-Tm-1)也与TM-4的横纹肌同工型一起在胚胎心脏中表达。在成年心脏中,表达的是TM-4,然而,α-Tm-1和α-Tm-2同工型的表达都大幅减少或下调。有趣的是,我们在胚胎和成年骨骼肌中未能检测到α-Tm-2的表达,但是α-Tm-1同工型在胚胎和成年骨骼肌中都有表达。对α-TM基因的其他可能同工型,即α-平滑肌原肌球蛋白(α-Sm)、α-成纤维细胞-1(α-F1)和α-成纤维细胞-2(α-F2)的检测显示,它们在胚胎心脏中有表达,并且在成年心脏中每种同工型都显著减少。为了阐明新发现的原肌球蛋白同工型在鸡中的作用,我们将α-Tm-1和α-Tm-2的绿色荧光蛋白融合蛋白分别异位表达到从新生大鼠分离的心肌细胞中。每种同工型都整合到有组织的肌原纤维中。我们的结果表明,在鸡心脏发育过程中,α-TM基因可能受到正、负转录调控。