Temperature sensitive point mutations in fission yeast tropomyosin have long range effects on the stability and function of the actin-tropomyosin copolymer.
Temperature sensitive point mutations in fission yeast tropomyosin have long range effects on the stability and function of the actin-tropomyosin copolymer.
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裂变酵母菌菌素中温度敏感的点突变对肌动蛋白 - 肌球蛋白共聚物的稳定性和功能具有长度的影响。
DOI：
10.1016/j.bbrc.2017.10.109
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发表时间：
2018-11-25
影响因子：
3.1
通讯作者：
Mulvihill DP
中科院分区：
文献类型：
--
作者：
Johnson CA;Brooker HR;Gyamfi I;O'Brien J;Ashley B;Brazier JE;Dean A;Embling J;Grimsey E;Tomlinson AC;Wilson EG;Geeves MA;Mulvihill DP
肌动蛋白的细胞骨架是由调节肌动蛋白结合蛋白，微调肌动蛋白聚合物的动态特性，以调节功能。一种这样的肌动蛋白结合蛋白是原肌球蛋白（Tpm），一种高度保守的α-螺旋二聚体，其稳定肌动蛋白并调节与其他蛋白质的相互作用。Tpm的温度敏感突变体是研究肌动蛋白丝依赖过程的宝贵工具，对细胞的生存力至关重要。在这里，我们调查的裂变酵母Tpm突变体的温度敏感性的分子基础，未能进行胞质分裂在限制性温度。收缩肌动球蛋白环（CAR）收缩以及细胞形状和大小的比较显示，cdc8.110或cdc8.27突变等位基因在细胞周期中的温度敏感性和对肌动蛋白依赖功能的影响方面显示出显着差异。体外分析显示，突变蛋白质显示出不同的热稳定性降低，并意外地产生两个离散的展开域乙酰化时，其氨基末端。我们的研究结果表明，如何微妙的结构变化（点突变或乙酰化）改变的稳定性，而不仅仅是这个保守的细胞骨架蛋白的离散区域，但整个分子。这差异性地影响这种必需的细胞骨架蛋白的稳定性和细胞组织。裂殖酵母温度敏感原肌球蛋白突变体的克隆、表达及特性分析。详细的体外分析温度对这些突变体的影响。比较体内突变对裂殖酵母内肌动蛋白环功能的影响。表明结构的细微变化改变了含原肌球蛋白聚合物的长期稳定性。
The actin cytoskeleton is modulated by regulatory actin-binding proteins which fine-tune the dynamic properties of the actin polymer to regulate function. One such actin-binding protein is tropomyosin (Tpm), a highly-conserved alpha-helical dimer which stabilises actin and regulates interactions with other proteins. Temperature sensitive mutants of Tpm are invaluable tools in the study of actin filament dependent processes, critical to the viability of a cell. Here we investigated the molecular basis of the temperature sensitivity of fission yeast Tpm mutants which fail to undergo cytokinesis at the restrictive temperatures. Comparison of Contractile Actomyosin Ring (CAR) constriction as well as cell shape and size revealed the cdc8.110 or cdc8.27 mutant alleles displayed significant differences in their temperature sensitivity and impact upon actin dependent functions during the cell cycle. In vitro analysis revealed the mutant proteins displayed a different reduction in thermostability, and unexpectedly yield two discrete unfolding domains when acetylated on their amino-termini. Our findings demonstrate how subtle changes in structure (point mutations or acetylation) alter the stability not simply of discrete regions of this conserved cytoskeletal protein but of the whole molecule. This differentially impacts the stability and cellular organisation of this essential cytoskeletal protein. Cloning, expression and characterisation of fission yeast temperature sensitive tropomyosin mutants. Detailed in vitro analysis on the impact of temperature upon these mutants. Comparison with in vivo impact of mutations upon actin ring function within the fission yeast. Demonstrates that subtle changes in structure alter the long range stability of Tropomyosin containing polymers.
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影响因子：
3.3
作者：
Clayton JE;Pollard LW;Sckolnick M;Bookwalter CS;Hodges AR;Trybus KM;Lord M
通讯作者：
Lord M
影响因子：
7.8
作者：
DREES, B;BROWN, C;BRETSCHER, A
通讯作者：
BRETSCHER, A
影响因子：
4
作者：
Gunning, Peter W.;Hardeman, Edna C.;Mulvihill, Daniel P.
通讯作者：
Mulvihill, Daniel P.
DOI：
10.1073/pnas.0509269102
发表时间：
2005-12-27
影响因子：
11.1
作者：
Brown, JH;Zhou, ZC;Cohen, C
通讯作者：
Cohen, C
影响因子：
3.3
作者：
Motegi, F;Arai, R;Mabuchi, I
通讯作者：
Mabuchi, I