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])
ISSN0076-6879
卷号658页码:83-109
发表状态已发表
DOI10.1016/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
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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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