Differential splicing of the large sarcomeric protein nebulin during skeletal muscle development
Differential splicing of the large sarcomeric protein nebulin during skeletal muscle development
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DOI：
10.1016/j.jsb.2010.02.014
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发表时间：
2010-05-01
影响因子：
3
通讯作者：
Granzier, Henk
中科院分区：
文献类型：
--
作者：
Buck, Danielle;Hudson, Bryan D.;Granzier, Henk
我们研究了星云蛋白的差异剪接，星云蛋白是一种在骨骼肌中发现的巨大丝状F-肌动蛋白结合蛋白（M-r类似于700-800 kDa）。Nebulin跨越细丝长度，其C-末端锚定在Z-盘中，并且其N-末端区域朝向细丝尖端定位。各种证据表明，星云蛋白在骨骼肌的细丝和Z盘结构中起重要作用。在目前的工作中，我们研究了在一系列的肌肉类型在出生后的发展，并进行转录本研究与小鼠nebulin外显子微阵列，我们开发的，其结果被证实了RT-PCR。我们还进行了蛋白质的研究，高分辨率SDS-琼脂糖凝胶和蛋白质印迹，和结构的研究，电子显微镜。我们发现在出生后的比目鱼肌发育过程中，在星云蛋白的超重复区域和Z盘区域的剪接发生了重大变化;有趣的是，这些变化在其他肌肉类型中是不存在的。三个新的Z盘外显子，以前描述的小鼠基因，在出生后的比目鱼肌发育过程中上调，这与Z盘宽度的显着增加。这些发现支持了星云蛋白在Z盘宽度调节中起重要作用的观点。总之，我们发现了星云蛋白的超级重复区域和Z盘区域的变化，这些变化是肌肉类型特异性的，并且它们与肌节结构的差异相关。(C)2010年爱思唯尔公司All rights reserved.
We studied differential splicing of nebulin, a giant filamentous F-actin binding protein (M-r similar to 700-800 kDa) that is found in skeletal muscle. Nebulin spans the thin filament length, its C-terminus is anchored in the Z-disc, and its N-terminal region is located toward the thin filament pointed end. Various lines of evidence indicate that nebulin plays important roles in thin filament and Z-disc structure in skeletal muscle. In the present work we studied nebulin in a range of muscle types during postnatal development and performed transcript studies with a mouse nebulin exon microarray, developed by us, whose results were confirmed by RT-PCR. We also performed protein studies with high-resolution SDS-agarose gels and Western blots, and structural studies with electron microscopy. We found during postnatal development of the soleus muscle major changes in splicing in both the super-repeat region and the Z-disc region of nebulin; interestingly, these changes were absent in other muscle types. Three novel Z-disc exons, previously described in the mouse gene, were upregulated during postnatal development of soleus muscle and this was correlated with a significant increase in Z-disc width. These findings support the view that nebulin plays an important role in Z-disc width regulation. In summary, we discovered changes in both the super-repeat region and the Z-disc region of nebulin, that these changes are muscle-type specific, and that they correlate with differences in sarcomere structure. (C) 2010 Elsevier Inc. All rights reserved.