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CTS tag-based methods for investigating mitochondrial RNA modification factors in Trypanosoma brucei | |
2021 | |
发表期刊 | METHODS IN ENZYMOLOGY (IF:1.682[JCR-2021],2.071[5-Year]) |
ISSN | 0076-6879 |
卷号 | 658页码:83-109 |
发表状态 | 已发表 |
DOI | 10.1016/bs.mie.2021.06.004 |
摘要 | Unicellular parasite Trypanosoma brucei maintains an elaborate mitochondrial mRNA processing pathway including 3'-5' exonucleolytic trimming of primary precursors, 5' and 3' modifications, and, in most cases, massive U-insertion/deletion editing. Whereas the role of editing in restoring protein coding sequence is apparent, recent developments suggest that terminal modifications are equally critical for generating a stable translationally competent messenger. The enzymatic activities responsible for 5' pyrophosphate hydrolysis, 3' adenylation and uridylation, and 3'-5' decay are positively and negatively regulated by pentatricopeptide repeat-containing (PPR) proteins. These sequence-specific RNA binding factors typically contain arrays of 35-amino acid repeats each of which recognizes a single nucleotide. Here, we introduce a combinatorial CTS affinity tag, which underlies a suite of methods for PPR proteins purification, in vivo RNA binding sites mapping and sub-cellular localization studies. These approaches should be applicable to most trypanosomal RNA binding proteins. |
收录类别 | SCIE |
WOS研究方向 | Biochemistry & Molecular Biology ; Biotechnology & Applied Microbiology |
WOS类目 | Biochemical Research Methods ; Biochemistry & Molecular Biology ; Biotechnology & Applied Microbiology |
WOS记录号 | WOS:000709781400006 |
出版者 | ELSEVIER ACADEMIC PRESS INC |
原始文献类型 | Review; Book Chapter |
引用统计 | 正在获取...
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文献类型 | 期刊论文 |
条目标识符 | https://kms.shanghaitech.edu.cn/handle/2MSLDSTB/128528 |
专题 | 生命科学与技术学院_PI研究组_张力烨组 生命科学与技术学院_本科生 |
通讯作者 | Aphasizheva, Inna |
作者单位 | 1.Boston Univ, Dept Mol & Cell Biol, Med Campus, Boston, MA 02215 USA; 2.ShanghaiTech Univ, Sch Life Sci & Technol, Shanghai, Peoples R China; 3.Boston Univ, Dept Biochem, Med Campus, Boston, MA 02215 USA |
推荐引用方式 GB/T 7714 | Aphasizheva, Inna,Suematsu, Takuma,Vacas, Andres,et al. CTS tag-based methods for investigating mitochondrial RNA modification factors in Trypanosoma brucei[J]. METHODS IN ENZYMOLOGY,2021,658:83-109. |
APA | Aphasizheva, Inna.,Suematsu, Takuma.,Vacas, Andres.,Wang, Hong.,Fan, Chenyu.,...&Aphasizhev, Ruslan.(2021).CTS tag-based methods for investigating mitochondrial RNA modification factors in Trypanosoma brucei.METHODS IN ENZYMOLOGY,658,83-109. |
MLA | Aphasizheva, Inna,et al."CTS tag-based methods for investigating mitochondrial RNA modification factors in Trypanosoma brucei".METHODS IN ENZYMOLOGY 658(2021):83-109. |
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