Structural and biochemical insights into the recognition of RNA helicase CGH-1 by CAR-1 in C. elegans
Structural and biochemical insights into the recognition of RNA helicase CGH-1 by CAR-1 in C. elegans
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DOI：
10.1016/j.bbrc.2021.02.119
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发表时间：
2021
期刊：
影响因子：
--
通讯作者：
Jingjun Hong
中科院分区：
文献类型：
--
作者：
Yong Zhang;Mengqi Lv;Fudong Li;Meili Li;Jiahai Zhang;Yunyu Shi;Jingjun Hong
A protein-RNA complex containing the RNA helicase CGH-1 and a germline specific RNA-binding protein.CAR-1 is involved in various aspects of function in C. elegans. However, the structural basis for the assembly.of this protein complex remains unclear. Here, we elucidate the molecular basis of the recognition.of CGH-1 by CAR-1. Additionally, we found that the ATPase activity of CGH-1 is stimulated by NTL-.1a MIF4G domain in vitro. Furthermore, we determined the structures of the two RecA-like domains of.CGH-1 by X-ray crystallography at resolutions of 1.85 and 2.40 Å, respectively. Structural and biochemical.approaches revealed a bipartite interface between CGH-1 RecA2 and the FDF-TFG motif of CAR-1. NMR.and structure-based mutations in CGH-1 RecA2 or CAR-1 attenuated or disrupted CGH-1 binding to CAR-.1, assessed by ITC and GST-pulldown in vitro. These findings provide insights into a conserved mechanism.in the recognition of CGH-1 by CAR-1. Together, our data provide the missing physical links in.understanding the assembly and function of CGH-1 and CAR-1 in C. elegans.
