Caldesmon is necessary for maintaining the actin and intermediate filaments in cultured bladder smooth muscle cells

被引:19
作者
Deng, Maoxian
Mohanan, Sunish
Polyak, Erzsebet
Chacko, Samuel
机构
[1] Univ Penn, Dept Surg, Div Surg, Glenolden, PA 19036 USA
[2] Univ Penn, Dept Pathol, Philadelphia, PA 19104 USA
来源
CELL MOTILITY AND THE CYTOSKELETON | 2007年 / 64卷 / 12期
关键词
caldesmon; actin; intermediate filament; smooth muscle cell;
D O I
10.1002/cm.20236
中图分类号
Q2 [细胞生物学];
学科分类号
071009 ; 090102 ;
摘要
Caldesmon (CaD), a component of microfilaments in all cells and thin filaments in smooth muscle cells, is known to bind to actin, tropomyosin, calmodulin, and myosin and to inhibit actin-activated ATP hydrolysis by smooth muscle myosin. Thus, it is believed to regulate smooth muscle contraction, cell motility and the cytoskeletal structure. Using bladder smooth muscle cell cultures and RNA interference (RNAi) technique, we show that the organization of actin into microfilaments in the cytoskeleton is diminished by siRNA-mediated CaD silencing. CaD silencing significantly decreased the amount of polymerized actin (F-actin), but the expression of actin was not altered. Additionally, we find that CaD is associated with 10 nm intermediatesized filaments (IF) and in vitro binding assay reveals that it binds to vimentin and desmin proteins. Assembly of vimentin and desmin into IF is also affected by CaD silencing, although their expression is not significantly altered when CaD is silenced. Electronmicroscopic analyses of the siRNA-treated cells showed the presence of myosin filaments and a few surrounding actin filaments, but the distribution of microfilament bundles was sparse. Interestingly, the decrease in CaD expression had no effect on tubulin expression and distribution of microtubules in these cells. These results demonstrate that CaD is necessary for the maintenance of actin microfilaments and intermediate-sized filaments in the cytoskeletal structure. This finding raises the possibility that the cytoskeletal structure in smooth muscle is affected when CaD expression is altered, as in smooth muscle de-differentiation and hypertrophy seen in certain pathological conditions.
引用
收藏
页码:951 / 965
页数:15
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