Biochemical and biophysical characterization of the deadenylase CrCaf1 from Chlamydomonas reinhardtii.
Biochemical and biophysical characterization of the deadenylase CrCaf1 from Chlamydomonas reinhardtii.
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莱茵衣藻脱腺苷酶 CrCaf1 的生化和生物物理表征
DOI：
10.1371/journal.pone.0069582
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发表时间：
2013
期刊：
影响因子：
3.7
通讯作者：
Yan YB
中科院分区：
文献类型：
--
作者：
Zhang JQ;He GJ;Yan YB
mRNA周转的调控越来越被认为是转录后水平基因表达调控的热点。在真核细胞中，大多数mRNA通过去腺苷酸化依赖性途径降解，其中去除poly（A）尾是初始和限速步骤。Caf 1是一种从3′-端特异性降解poly（A）的去腺苷酶，从酵母到酵母高度保守。高等植物中的Caf 1 s已被证明参与植物发育和胁迫反应。然而，对植物中Caf 1 s的生物化学和生物物理特性知之甚少。本研究克隆了莱茵衣藻crcaf 1基因，并对重组蛋白的性质进行了研究。结果表明，CrCaf 1是一种去腺苷酶，具有Caf 1家族保守的序列基序、结构特征和催化特性。CrCaf 1以分布方式降解poly（A），最佳反应条件为pH 7和35°C。CrCaf 1与Mg ~（2+）、Mn ~（2+）配位时活性相近，而与Ca ~（2+）、Zn ~（2+）配位时几乎失活。Zn 2+可诱导CrCaf 1聚集并破坏其天然结构，而Mg 2+、Mn 2+和Ca 2+可通过减少蛋白聚集来稳定CrCaf 1，使其不受热变性的影响。在各种金属离子中，Mn 2+对CrCaf 1稳定性的保护作用最强，提示Mn 2+可能在调控CrCaf 1稳定性中发挥作用。reinhardtii细胞在某些应激条件下的生长。这些发现为进一步研究CrCaf 1在C.莱因哈德氏菌
The modulation of mRNA turnover has been increasingly recognized as a hotpoint for gene expression regulation at the post-transcriptional level. In eukaryotic cells, most mRNAs are degraded via the deadenylation-dependent pathway, in which the removal of the poly(A) tail is the initial and rate-limiting step. Caf1, a deadenylase specifically degrades poly(A) from the 3′-end, is highly conserved from yeast to mammalians. Caf1s in higher plants have been shown to be involved in plant development and stress response. However, little is known about the biochemical and biophysical properties of Caf1s in plants. In this research, we cloned the crcaf1 gene from Chlamydomonas reinhardtii and studied the properties of the recombinant proteins. The results showed that CrCaf1 was a deadenylase with conserved sequence motifs, structural features, and catalytic properties of the Caf1 family. CrCaf1 degraded poly(A) in a distributive mode with the optimal reacting conditions at pH 7 and 35°C. CrCaf1 had similar activity when coordinated with Mg2+ and Mn2+, while the enzyme bound to Ca2+ or Zn2+ was almost inactivated. Zn2+ could induce CrCaf1 aggregation with the disruption of the native structure, while Mg2+, Mn2+ and Ca2+ could stabilize CrCaf1 against thermal denaturation by reducing protein aggregation. Among the various metal ions, Mn2+ showed the strongest protective effect on CrCaf1 stability, implying that Mn2+ might play a role in regulating CrCaf1 stability in the C. reinhardtii cells under some stressed conditions. These findings provide a starting point for further investigation of the physiological functions of CrCaf1 in C. reinhardtii.
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影响因子：
14.9
作者：
Ohn T;Chiang YC;Lee DJ;Yao G;Zhang C;Denis CL
通讯作者：
Denis CL
四个保守酸性残基在聚 (A) 特异性核糖核酸酶热稳定性中的不同作用
影响因子：
5.6
作者：
He GJ;Liu WF;Yan YB
通讯作者：
Yan YB
影响因子：
16
作者：
Reinhardt HC;Hasskamp P;Schmedding I;Morandell S;van Vugt MA;Wang X;Linding R;Ong SE;Weaver D;Carr SA;Yaffe MB
通讯作者：
Yaffe MB
N 末端调节人 Caf1 活性、结构稳定性和聚集
DOI：
10.1016/j.ijbiomac.2012.05.032
发表时间：
2012-11-01
影响因子：
8.2
作者：
Feng, Li-Kui;Yan, Yong-Bin
通讯作者：
Yan, Yong-Bin
影响因子：
5.3
作者：
Gera, JF;Baker, EJ
通讯作者：
Baker, EJ