The Human CCHC-type Zinc Finger Nucleic Acid-Binding Protein Binds G-Rich Elements in Target mRNA Coding Sequences and Promotes Translation.
The Human CCHC-type Zinc Finger Nucleic Acid-Binding Protein Binds G-Rich Elements in Target mRNA Coding Sequences and Promotes Translation.
复制标题
DOI：
10.1016/j.celrep.2017.02.080
复制
发表时间：
2017-03-21
期刊：
影响因子：
8.8
通讯作者：
Juranek SA
中科院分区：
文献类型：
--
作者：
Benhalevy D;Gupta SK;Danan CH;Ghosal S;Sun HW;Kazemier HG;Paeschke K;Hafner M;Juranek SA
CCHC型锌指核酸结合蛋白（CNBP/ZNF 9）是真核生物中保守的蛋白，对哺乳动物胚胎发育至关重要。它已被牵连在转录以及转录后的基因调控，然而，其核酸配体和分子功能仍然难以捉摸。在这里，我们使用多个系统范围的方法来识别CNBP的目标和功能。我们使用光活化核糖核苷增强的交联和免疫沉淀（PAR-CLIP），以确定8420 CNBP结合位点的4178 mRNA。CNBP优先结合靶mRNA编码序列中的富含G的元件，其中大部分先前被发现在体外形成G-四链体和其他稳定结构。功能分析，包括RNA测序，核糖体分析和定量质谱分析，显示CNBP结合不影响靶mRNA丰度，而是增加其翻译效率。考虑到CNBP结合阻止了体外G四链体结构的形成，我们假设CNBP通过解析mRNA上的稳定结构来支持翻译。Benhalevy等人使用全系统方法表征RNA结合蛋白CNBP/ZNF 9。他们发现CNBP优先结合在先前发现的在体外形成G四链体和其他结构的mRNA区域。核糖体分析表明，CNBP增强了这些位点的翻译，这可能会形成核糖体的路障。
The CCHC-type Zinc Finger Nucleic Acid Binding Protein (CNBP/ZNF9) is conserved in eukaryotes and essential for embryonic development in mammals. It has been implicated in transcriptional as well as post-transcriptional gene regulation; however, its nucleic acid ligands and molecular function remain elusive. Here, we use multiple systems-wide approaches to identify CNBP targets and function. We used Photoactivatable Ribonucleoside Enhanced Crosslinking and Immunoprecipitation (PAR-CLIP) to identify 8420 CNBP binding sites on 4178 mRNAs. CNBP preferentially bound G-rich elements in the target mRNA coding sequences, most of which were previously found to form G-quadruplex and other stable structures in vitro. Functional analyses, including RNA sequencing, ribosome profiling, and quantitative mass spectrometry, revealed that CNBP binding did not influence target mRNA abundance but rather increased their translational efficiency. Considering that CNBP binding prevented G quadruplex structure formation in vitro, we hypothesize that CNBP is supporting translation by resolving stable structures on mRNAs. Benhalevy et al. characterize the RNA binding protein CNBP/ZNF9 using systems-wide approaches. They find that CNBP preferentially binds at mRNA regions previously found to form G quadruplex and other structures in vitro. Ribosome profiling revealed that CNBP enhances translation across these sites, which potentially form roadblocks for the ribosome.