Substrate specificity of human MCPIP1 endoribonuclease.
Substrate specificity of human MCPIP1 endoribonuclease.
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DOI：
10.1038/s41598-018-25765-2
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发表时间：
2018-05-09
影响因子：
4.6
通讯作者：
Jura J
中科院分区：
文献类型：
--
作者：
Wilamowski M;Gorecki A;Dziedzicka-Wasylewska M;Jura J
MCPIP1，也被称为regase -1，是一种核糖核酸酶，对调节炎症过程相关转录本的稳定性至关重要。在这里，我们报道MCPIP1通过降解几种茎环RNA结构和单链RNA作为一种内切酶。我们的研究发现，在白介素-6转录物的3 '非翻译区衍生的茎环中存在裂解位点。此外，MCPIP1诱导茎环结构环基序的内切酶裂解。此外，我们观察到MCPIP1可以切割单链RNA片段。然而，短于6个核苷酸的RNA底物不受MCPIP1溶核活性的进一步影响。在本研究中，我们还测定了全长MCPIP1D141N及其核糖核酸酶结构域PIN D141N与12个寡核苷酸底物的解离常数。MCPIP1D141N与RNA靶点的平衡结合常数（Kd）约为10 nM。有趣的是，我们观察到PIN结构域中锌指的存在增加了该蛋白片段对25个核苷酸长的茎环RNA的亲和力，而不是对较短的RNA的亲和力。此外，MCPIP1和PIN蛋白的大小排斥层析表明，MCPIP1在与RNA底物相互作用时发生了同质寡聚化。我们的研究结果深入了解了MCPIP1底物识别的机制及其对各种寡核苷酸的亲和力。
MCPIP1, also known as Regnase-1, is a ribonuclease crucial for regulation of stability of transcripts related to inflammatory processes. Here, we report that MCPIP1 acts as an endonuclease by degrading several stem-loop RNA structures and single-stranded RNAs. Our studies revealed cleavage sites present in the stem-loops derived from the 3′ untranslated region of the interleukin-6 transcript. Furthermore, MCPIP1 induced endonuclease cleavage at the loop motif of stem-loop structures. Additionally, we observed that MCPIP1 could cleave single-stranded RNA fragments. However, RNA substrates shorter than 6 nucleotides were not further affected by MCPIP1 nucleolytic activity. In this study, we also determined the dissociation constants of full-length MCPIP1D141N and its ribonuclease domain PIN D141N with twelve oligonucleotides substrates. The equilibrium binding constants (Kd) for MCPIP1D141N and the RNA targets were approximately 10 nM. Interestingly, we observed that the presence of a zinc finger in the PIN domain increases the affinity of this protein fragment to 25-nucleotide-long stem-loop RNA but not to shorter ones. Furthermore, size exclusion chromatography of the MCPIP1 and PIN proteins suggested that MCPIP1 undergoes homooligomerization during interaction with RNA substrates. Our results provide insight into the mechanism of MCPIP1 substrate recognition and its affinity towards various oligonucleotides.
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影响因子：
2.9
作者：
Salvi, G;De los Rios, P;Vendruscolo, M
通讯作者：
Vendruscolo, M
DOI：
10.4049/jimmunol.1601551
发表时间：
2017-01-15
期刊：
Journal of immunology (Baltimore, Md. : 1950)
影响因子：
--
作者：
Monin L;Gudjonsson JE;Childs EE;Amatya N;Xing X;Verma AH;Coleman BM;Garg AV;Killeen M;Mathers A;Ward NL;Gaffen SL
通讯作者：
Gaffen SL
影响因子：
11.2
作者：
Lu W;Ning H;Gu L;Peng H;Wang Q;Hou R;Fu M;Hoft DF;Liu J
通讯作者：
Liu J
影响因子：
3.7
作者：
Lipert B;Wilamowski M;Gorecki A;Jura J
通讯作者：
Jura J
基于多基因的异翅目（昆虫纲：半翅目）高级系统发育和首次分化时间估计
影响因子：
3.7
作者：
Li M;Tian Y;Zhao Y;Bu W
通讯作者：
Bu W