Unveiling the Essential Role of Arkadia's Non-RING Elements in the Ubiquitination Process.
Unveiling the Essential Role of Arkadia's Non-RING Elements in the Ubiquitination Process.
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DOI：
10.3390/ijms231810585
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发表时间：
2022-09-13
影响因子：
5.6
通讯作者：
Spyroulias, Georgios A.
中科院分区：
文献类型：
--
作者：
Birkou, Maria;Delegkou, Georgia N.;Marousis, Konstantinos D.;Fragkaki, Nefeli;Toro, Tamara;Episkopou, Vasso;Spyroulias, Georgios A.
Arkadia是TGFβ-SMAD 2/3通路的正调节因子，通过其C-末端RING-H2结构域发挥作用，并靶向降解其负调节因子。在这里，我们探讨的作用以外的区域的RING域（非RING元件）的Arkadia的E2-E3相互作用。使用Arkadia RING 68 aa和Arkadia 119 aa多肽解决非RING元件的贡献。在119个氨基酸的Arkadia多肽中，高度保守的NRGA（天冬酰胺-精氨酸-甘氨酸-丙氨酸）和TIER（苏氨酸-异亮氨酸-谷氨酰胺-精氨酸）基序已被证明是pSMAD 2/3底物识别和体内泛素化所需的。然而，NRGA和TIER基序在Arkadia的酶活性中的作用尚未得到解决。在这里，使用E2酶、UBCH 5 B、C85 S UBCH 5 B-Ub氧酯水解和自动泛素化测定的核磁共振相互作用研究来解决非RING元件在E2-E3相互作用和RING的酶活性中的作用。这些结果支持包括NRGA和TIER基序在内的非RING元件是E2-E3识别和相互作用以及有效的自动泛素化所必需的。此外，虽然已知Arkadia同种型-2及其同源物Arkadia 2C与游离泛素相互作用，但此处的结果表明Arkadia同种型-1不与游离泛素相互作用。
Arkadia is a positive regulator of the TGFβ-SMAD2/3 pathway, acting through its C-terminal RING-H2 domain and targeting for degradation of its negative regulators. Here we explore the role of regions outside the RING domain (non-RING elements) of Arkadia on the E2-E3 interaction. The contribution of the non-RING elements was addressed using Arkadia RING 68 aa and Arkadia 119 aa polypeptides. The highly conserved NRGA (asparagine-arginine-glycine-alanine) and TIER (threonine-isoleucine-glutamine-arginine) motifs within the 119 aa Arkadia polypeptide, have been shown to be required for pSMAD2/3 substrate recognition and ubiquitination in vivo. However, the role of the NRGA and TIER motifs in the enzymatic activity of Arkadia has not been addressed. Here, nuclear magnetic resonance interaction studies with the E2 enzyme, UBCH5B, C85S UBCH5B-Ub oxyester hydrolysis, and auto-ubiquitination assays were used to address the role of the non-RING elements in E2-E3 interaction and in the enzymatic activity of the RING. The results support that the non-RING elements including the NRGA and TIER motifs are required for E2-E3 recognition and interaction and for efficient auto-ubiquitination. Furthermore, while Arkadia isoform-2 and its close homologue Arkadia 2C are known to interact with free ubiquitin, the results here showed that Arkadia isoform-1 does not interact with free ubiquitin.
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影响因子：
64.8
作者：
Jumper J;Evans R;Pritzel A;Green T;Figurnov M;Ronneberger O;Tunyasuvunakool K;Bates R;Žídek A;Potapenko A;Bridgland A;Meyer C;Kohl SAA;Ballard AJ;Cowie A;Romera-Paredes B;Nikolov S;Jain R;Adler J;Back T;Petersen S;Reiman D;Clancy E;Zielinski M;Steinegger M;Pacholska M;Berghammer T;Bodenstein S;Silver D;Vinyals O;Senior AW;Kavukcuoglu K;Kohli P;Hassabis D
通讯作者：
Hassabis D
影响因子：
16
作者：
Das, Ranabir;Mariano, Jennifer;Tsai, Yien Che;Kalathur, Ravi C.;Kostova, Zlatka;Li, Jess;Tarasov, Sergey G.;McFeeters, Robert L.;Altieri, Amanda S.;Ji, Xinhua;Byrd, R. Andrew;Weissman, Allan M.
通讯作者：
Weissman, Allan M.
影响因子：
11.2
作者：
Le Scolan, Erwan;Zhu, Qingwei;Luo, Kunxin
通讯作者：
Luo, Kunxin
影响因子：
14.8
作者：
Chaugule, Viduth K.;Arkinson, Connor;Walden, Helen
通讯作者：
Walden, Helen
影响因子：
4.8
作者：
Li, Shengjian;Liang, Yu-He;Weissman, Allan M.
通讯作者：
Weissman, Allan M.