Actin isoform utilization during differentiation and remodeling of BC3H1 myogenic cells

Actin isoform utilization during differentiation and remodeling of BC3H1 myogenic cells
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BC3H1 肌原细胞分化和重塑过程中肌动蛋白异构体的利用

DOI:
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发表时间:
1997
影响因子:
4
通讯作者:
A. Strauch
A. Strauch
中科院分区:
生物学2区
文献类型:
--
作者:
Guang Qu;Huara Yan;A. Strauch

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利用小鼠成肌细胞和一种双功能化学交联剂,研究了新合成的血管平滑肌(α-肌动蛋白)和非肌肉(β-和γ-肌动蛋白)肌动蛋白单体在肌发生过程中聚合成天然F-肌动蛋白细丝结构的情况。通过对BC3H1成肌细胞和肌细胞的苯双马来酰亚胺交联部分的SDS-PAGE分析,鉴定出两种肌动蛋白二聚体。P-二聚体来源于富含F-肌动蛋白的不溶于洗涤剂的细胞骨架。脉冲追逐分析表明,D-二聚体最初与细胞骨架有关,但随后在含有非丝状或聚集的肌动蛋白池的溶解肌肉细胞中积累。免疫印迹分析表明,非肌肉肌动蛋白和平滑肌肌动蛋白都能形成这两种类型的二聚体。然而,在发育中的成肌细胞中,平滑肌α-肌动蛋白的诱导与D-二聚体水平的增加相一致,这可能有助于肌动蛋白应激纤维的组装。平滑肌α-肌动蛋白被迅速用于分化成肌细胞,在细胞骨架中组装耐提取的F-肌动蛋白细丝,而非肌肉的β-和γ-肌动蛋白细丝更容易被含有三磷酸腺苷和EGTA的提取缓冲液从细胞骨架中解离。这些数据表明,在发育中的BC3H1肌源性细胞中,细胞结构重构伴随着肌动蛋白异构体的选择性利用,这种利用有效地将多个异肌动蛋白分离到不同的亚细胞域和/或超分子实体。J.细胞。生物化学。67:514-527,1997。©1997 Wiley-Liss,Inc.
Mouse BC3H1 myogenic cells and a bi‐functional chemical cross linking reagent were utilized to investigate the polymerization of newly‐synthesized vascular smooth muscle (α‐actin) and non‐muscle (β‐ and γ‐actin) actin monomers into native F‐actin filament structures during myogenesis. Two actin dimer species were identified by SDS‐PAGE analysis of phenylenebismaleimide‐cross linked fractions of BC3H1 myoblasts and myocytes. P‐dimer was derived from the F‐actin‐enriched, detergent‐insoluble cytoskeleton. Pulse‐chase analysis revealed that D‐dimer initially was associated with the cytoskeleton but then accumulated in the soluble fraction of lysed muscle cells that contained a non‐filamentous or aggregated actin pool. Immunoblot analysis indicated that non‐muscle and smooth muscle actins were capable of forming both types of dimer. However, induction of smooth muscle α‐actin in developing myoblasts coincided with an increase in D‐dimer level which may facilitate actin stress fiber assembly. Smooth muscle α‐actin was rapidly utilized in differentiating myoblasts to assemble extraction‐resistant F‐actin filaments in the cytoskeleton whereas non‐muscle β‐ and γ‐actin filaments were more readily dissociated from the cytoskeleton by an extraction buffer containing ATP and EGTA. The data indicate that cytoarchitectural remodeling in developing BC3H1 myogenic cells is accompanied by selective actin isoform utilization that effectively segregates multiple isoactins into different sub‐cellular domains and/or supramolecular entities. J. Cell. Biochem. 67:514–527, 1997. © 1997 Wiley‐Liss, Inc.
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