Mechanism of action of ABCF ATPases during translation

ABCF ATP酶在翻译过程中的作用机制

• 批准号: 398213262
• 负责人: Professor Dr. Daniel Nicodemus Wilson
• 金额: --
• 依托单位: Institut für Biochemie und Molekularbiologie
• 依托单位国家: 德国
• 项目类别: Research Grants
• 财政年份: 2018
• 资助国家: 德国
• 起止时间: 2017-12-31 至 2021-12-31
• 项目状态: 已结题
• 来源: https://gepris.dfg.de/gepris/projekt/398213262?language=en
• 关键词: Mechanism; action; ABCF; ATPases; during

In cells, the synthesis of proteins occurs on ribosomes and is facilitated by a plethora of specialized translation factors. In addition to translational GTPases, there is an emerging role for regulation of translation via translational ATPases, such as the elongation factor EF3 in eukaryotes and the antibiotic resistance (ARE) ABCF ATPases in bacteria. In eukaryotes, EF3 interacts with the ribosome during translation elongation, where it promotes release of the E-tRNA. There are also reports that under some conditions eEF3 can also promote ribosomal splitting. Only a single low (9.9 Å) resolution structure of an in vitro reconstituted yeast EF3-80S complex has been reported from over 10 years ago. Additionally, Yeast and fungi have many homologues of eEF3, such as New1p, however, it remains unclear whether they are also involved in translation. Here we propose to determine structures of EF3 and New1p in complex with the 80S ribosome using both in vitro and in vivo approaches. Such investigations will be necessary to understand why yeast and fungi require this factor for survival, and whether this function is taken over by another factor in higher eukaryotes. The ARE-ABCF ATPases are found in Gram-positive antibiotic-producing bacteria, such as Bacillus subtilis, as well as in pathogenic bacteria, such as Staphylococcus, Streptococcus, and Enterococcus. The ARE-ABCF proteins confer resistance to antibiotics that bind at or near the peptidyl-transferase center (PTC) of the ribosome, with different ARE-ABCF proteins having different antibiotic specificities. To date, there are no structures of the ARE-ABCF subfamily of proteins in complex with the ribosome, therefore, it remains unclear how these proteins bind to the ribosome and mechanistically how they confer resistance to the specific ribosome-targeting antibiotic classes. Here we propose to determine structures of ARE-ABCF proteins in complex with the ribosome to understand how these proteins bind to the ribosome and mechanistically how they confer resistance to the specific ribosome-targeting antibiotic classes. Given the ever-increasing emergence of multi drug resistant bacteria, understanding these novel bacterial resistance mechanisms will be important for development of new improved antibiotics.

在细胞中，蛋白质的合成发生在核糖体上，并由大量专门的翻译因子促进。除了翻译GTPases，还有一个通过翻译ATPases调节翻译的新角色，如真核生物中的延伸因子EF3和细菌中的抗生素抗性（ARE） ABCF ATPases。在真核生物中，EF3在翻译延伸过程中与核糖体相互作用，促进E-tRNA的释放。也有报道称，在某些情况下，eEF3也能促进核糖体分裂。只有一个低分辨率（9.9 Å）结构的体外重组酵母EF3-80S复合体已经报道了超过10年前。此外，酵母和真菌也有许多eEF3的同源物，如New1p，然而，它们是否也参与翻译尚不清楚。在这里，我们提出用体外和体内两种方法来确定EF3和New1p与80S核糖体复合物的结构。这样的研究对于理解为什么酵母和真菌需要这种因子来生存，以及这种功能是否在高等真核生物中被另一种因子所接管是必要的。are - abcf atp酶存在于革兰氏阳性抗生素产生细菌中，如枯草芽孢杆菌，以及致病菌中，如葡萄球菌、链球菌和肠球菌。ARE-ABCF蛋白赋予对结合在核糖体肽基转移酶中心（PTC）或其附近的抗生素的抗性，不同的ARE-ABCF蛋白具有不同的抗生素特异性。迄今为止，还没有发现are - abcf亚家族蛋白与核糖体复合物的结构，因此，尚不清楚这些蛋白如何与核糖体结合，以及它们如何从机制上赋予对特定核糖体靶向抗生素类的抗性。在这里，我们建议确定ARE-ABCF蛋白与核糖体复合物的结构，以了解这些蛋白如何与核糖体结合，以及它们如何从机制上赋予对特定核糖体靶向抗生素类的抗性。鉴于多重耐药细菌的不断出现，了解这些新的细菌耐药机制将对开发新的改良抗生素具有重要意义。

项目成果

A role for the Saccharomyces cerevisiae ABCF protein New1 in translation termination/recycling

• DOI: 10.1093/nar/gkz600
• 发表时间: 2019-09-19
• 期刊: NUCLEIC ACIDS RESEARCH
• 影响因子: 14.9
• 作者: Kasari, Villu;Pochopien, Agnieszka A.;Hauryliuk, Vasili
• 通讯作者: Hauryliuk, Vasili

Yeast translation elongation factor eEF3 promotes late stages of tRNA translocation.

• DOI: 10.15252/embj.2020106449
• 发表时间: 2021-03-15
• 期刊: The EMBO journal
• 作者: Ranjan N;Pochopien AA;Chih-Chien Wu C;Beckert B;Blanchet S;Green R;V Rodnina M;Wilson DN
• 通讯作者: Wilson DN