Pentatricopeptide repeat proteins stimulate mRNA adenylation/uridylation to activate mitochondrial translation in trypanosomes.
Pentatricopeptide repeat proteins stimulate mRNA adenylation/uridylation to activate mitochondrial translation in trypanosomes.
复制标题
DOI：
10.1016/j.molcel.2011.02.021
复制
发表时间：
2011-04-08
期刊：
影响因子：
16
通讯作者：
Aphasizhev R
中科院分区：
文献类型：
--
作者：
Aphasizheva I;Maslov D;Wang X;Huang L;Aphasizhev R
大多数锥虫线粒体前 mRNA 都会经历大量尿苷插入/删除编辑，从而创建开放阅读框。尽管已知在编辑前添加短 3' A 尾可以在编辑期间和编辑后稳定转录本，但将完全编辑的 mRNA 进行翻译的处理事件仍然未知。在这里，我们展示了含有五肽重复序列（PPR）蛋白的异二聚体，称为动质体聚腺苷酸化/尿苷酸化因子（KPAF）1和2，通过KPAP1聚（A）聚合酶和RET1末端尿苷基转移酶诱导A/U-杂聚物的编辑后添加。带有 200-300 个核苷酸长的 A/U 尾（而非短 A 尾）的编辑转录本在翻译核糖体复合物和亲和纯化的核糖体颗粒方面得到富集。 KPAF1 抑制导致 A/U 尾 mRNA 选择性丢失，并同时抑制蛋白质合成。这些结果将 A/U 延伸确立为具有翻译能力的 mRNA 的定义顺式元件。此外，我们证明 A/U 尾 mRNA 优先与小核糖体亚基相互作用，而编辑的底物和复合物与大亚基结合。
The majority of trypanosomal mitochondrial pre-mRNAs undergo massive uridine insertion/deletion editing which creates open reading frames. Although the pre-editing addition of short 3′ A-tails is known to stabilize transcripts during and after the editing, the processing event committing the fully-edited mRNAs to translation remained unknown. Here we show that a heterodimer of pentatricopeptide repeat-containing (PPR) proteins, termed kinetoplast polyadenylation/ uridylation factors (KPAFs) 1 and 2, induces the post-editing addition of A/U-heteropolymers by KPAP1 poly(A) polymerase and RET1 terminal uridyltransferase. Edited transcripts bearing 200–300 nucleotide-long A/U-tails, but not short A-tails, were enriched in translating ribosomal complexes and affinity-purified ribosomal particles. KPAF1 repression led to a selective loss of A/U-tailed mRNAs and concomitant inhibition of protein synthesis. These results establish A/U extensions as the defining cis-elements of translation-competent mRNAs. Furthermore, we demonstrate that A/U-tailed mRNA preferentially interacts with the small ribosomal subunit, whereas edited substrates and complexes bind to the large subunit.
登录
查看更多内容
影响因子：
64.5
作者：
Aphasizhev, R;Sbicego, S;Simpson, L
通讯作者：
Simpson, L
影响因子：
56.9
作者：
Schnaufer, A;Panigrahi, AK;Stuart, K
通讯作者：
Stuart, K
影响因子：
2.9
作者：
CHOMCZYNSKI, P;SACCHI, N
通讯作者：
SACCHI, N
影响因子：
64.5
作者：
Seiwert, SD;Heidmann, S;Stuart, K
通讯作者：
Stuart, K
影响因子：
11.4
作者：
Etheridge, Ronald D.;Aphasizheva, Inna;Aphasizhev, Ruslan
通讯作者：
Aphasizhev, Ruslan