Department of Cell and Developmental Biology, 107 Weiskotten Hall, State University of New York Upstate Medical University, 766 Irving Avenue, Syracuse, NY, 13210, USA.
Exp Cell Res. 2021 Jan 1;398(1):112388. doi: 10.1016/j.yexcr.2020.112388. Epub 2020 Nov 20.
Previous work with cultured cells has shown transcription of muscle genes by serum response factor (SRF) can be stimulated by actin polymerization driven by proteins of the formin family. However, it is not clear if endogenous formins similarly promote SRF-dependent transcription during muscle development in vivo. We tested whether formin activity promotes SRF-dependent transcription in striated muscle in the simple animal model, Caenorhabditis elegans. Our lab has shown FHOD-1 is the only formin that directly promotes sarcomere formation in the worm's striated muscle. We show here FHOD-1 and SRF homolog UNC-120 both support muscle growth and also muscle myosin II heavy chain A expression. However, while a hypomorphic unc-120 allele blunts expression of a set of striated muscle genes, these genes are largely upregulated or unchanged by absence of FHOD-1. Instead, pharmacological inhibition of the proteasome restores myosin protein levels in worms lacking FHOD-1, suggesting elevated proteolysis accounts for their myosin deficit. Interestingly, proteasome inhibition does not restore normal muscle growth to fhod-1(Δ) mutants, suggesting formin contributes to muscle growth by some alternative mechanism. Overall, we find SRF does not depend on formin to promote muscle gene transcription in a simple in vivo system.
先前的细胞培养实验表明,肌球蛋白基因的转录可以被肌动蛋白聚合所激活,而肌动蛋白聚合是由形成蛋白家族的蛋白所驱动的。然而,在体内肌肉发育过程中,内源性形成蛋白是否同样促进了 SRF 依赖性转录,目前还不清楚。我们在简单的动物模型秀丽隐杆线虫中,检测了形成蛋白活性是否能促进肌肉中 SRF 依赖性转录。我们的实验室已经表明,FHOD-1 是唯一一种能直接促进线虫有条纹肌肉肌节形成的形成蛋白。我们在这里表明,FHOD-1 和 SRF 同源物 UNC-120 都支持肌肉生长,也支持肌肉肌球蛋白 II 重链 A 的表达。然而,虽然 UNC-120 的一个弱等位基因削弱了一组有条纹肌肉基因的表达,但这些基因在 FHOD-1 缺失时,大部分被上调或没有变化。相反,蛋白酶体的药理学抑制在缺乏 FHOD-1 的蠕虫中恢复了肌球蛋白蛋白水平,这表明蛋白酶体的过度激活导致了它们的肌球蛋白缺乏。有趣的是,蛋白酶体抑制不能使 fhod-1(Δ)突变体恢复正常的肌肉生长,这表明形成蛋白通过某种替代机制促进肌肉生长。总的来说,我们发现 SRF 不需要形成蛋白来促进简单的体内系统中的肌肉基因转录。