دورية أكاديمية
HuR-miRNA complex activates RAS GTPase RalA to facilitate endosome targeting and extracellular export of miRNAs.
| العنوان: | HuR-miRNA complex activates RAS GTPase RalA to facilitate endosome targeting and extracellular export of miRNAs. |
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| المؤلفون: | Ghosh S; RNA Biology Research Laboratory, Molecular Genetics Division, CSIR-Indian Institute of Chemical Biology, Kolkata, India., Hom Choudhury S; RNA Biology Research Laboratory, Molecular Genetics Division, CSIR-Indian Institute of Chemical Biology, Kolkata, India., Mukherjee K; RNA Biology Research Laboratory, Molecular Genetics Division, CSIR-Indian Institute of Chemical Biology, Kolkata, India; Department of Pharmacology and Experimental Neuroscience, University of Nebraska Medical Center (UNMC), Nebraska, USA. Electronic address: kmukherjee@unmc.edu., Bhattacharyya SN; RNA Biology Research Laboratory, Molecular Genetics Division, CSIR-Indian Institute of Chemical Biology, Kolkata, India; Department of Pharmacology and Experimental Neuroscience, University of Nebraska Medical Center (UNMC), Nebraska, USA. Electronic address: sbhattacharyya@unmc.edu. |
| المصدر: | The Journal of biological chemistry [J Biol Chem] 2024 Mar; Vol. 300 (3), pp. 105750. Date of Electronic Publication: 2024 Feb 14. |
| نوع المنشور: | Journal Article |
| اللغة: | English |
| بيانات الدورية: | Publisher: Elsevier Inc. on behalf of American Society for Biochemistry and Molecular Biology Country of Publication: United States NLM ID: 2985121R Publication Model: Print-Electronic Cited Medium: Internet ISSN: 1083-351X (Electronic) Linking ISSN: 00219258 NLM ISO Abbreviation: J Biol Chem Subsets: MEDLINE |
| أسماء مطبوعة: | Publication: 2021- : [New York, NY] : Elsevier Inc. on behalf of American Society for Biochemistry and Molecular Biology Original Publication: Baltimore, MD : American Society for Biochemistry and Molecular Biology |
| مواضيع طبية MeSH: | Endosomes*/metabolism , MicroRNAs*/genetics , MicroRNAs*/metabolism , ral GTP-Binding Proteins*/metabolism , ELAV-Like Protein 1*/metabolism , Macrophages*/metabolism, Adenosine Triphosphate/metabolism ; rab5 GTP-Binding Proteins/genetics ; rab5 GTP-Binding Proteins/metabolism ; Humans ; HEK293 Cells |
| مستخلص: | Extracellular vesicles-mediated exchange of miRNA cargos between diverse types of mammalian cells is a major mechanism of controlling cellular miRNA levels and activity, thus regulating the expression of miRNA-target genes in both donor and recipient cells. Despite tremendous excitement related to extracellular vesicles-associated miRNAs as biomarkers or having therapeutic potential, the mechanism of selective packaging of miRNAs into endosomes and multivesicular bodies for subsequent extracellular export is poorly studied due to the lack of an in vitro assay system. Here, we have developed an in vitro assay with endosomes isolated from mammalian macrophage cells to follow miRNA packaging into endocytic organelles. The synthetic miRNAs, used in the assay, get imported inside the isolated endosomes during the in vitro reaction and become protected from RNase in a time- and concentration-dependent manner. The selective miRNA accumulation inside endosomes requires both ATP and GTP hydrolysis and the miRNA-binding protein HuR. The HuR-miRNA complex binds and stimulates the endosomal RalA GTPase to facilitate the import of miRNAs into endosomes and their subsequent export as part of the extracellular vesicles. The endosomal targeting of miRNAs is also very much dependent on the endosome maturation process that is controlled by Rab5 protein and ATP. In summary, we provide an in vitro method to aid in the investigation of the mechanism of miRNA packaging process for its export from mammalian macrophage cells. Competing Interests: Conflict of interest The authors declare that they have no conflicts of interest with the contents of this article. (Copyright © 2024 The Authors. Published by Elsevier Inc. All rights reserved.) |
| فهرسة مساهمة: | Keywords: HuR; RNA cargo trafficking; RNA import; Rab5; RalA GTPase; endosome isolation; endosomes; extracellular vesicles; miRNA |
| المشرفين على المادة: | 8L70Q75FXE (Adenosine Triphosphate) 0 (MicroRNAs) EC 3.6.5.2 (rab5 GTP-Binding Proteins) EC 3.6.1.- (RALA protein, human) EC 3.6.5.2 (ral GTP-Binding Proteins) 0 (ELAVL1 protein, human) 0 (ELAV-Like Protein 1) |
| تواريخ الأحداث: | Date Created: 20240215 Date Completed: 20240403 Latest Revision: 20240416 |
| رمز التحديث: | 20240417 |
| مُعرف محوري في PubMed: | PMC10956062 |
| DOI: | 10.1016/j.jbc.2024.105750 |
| PMID: | 38360271 |
| قاعدة البيانات: | MEDLINE |
Full Text Finder | تدمد: | 1083-351X |
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| DOI: | 10.1016/j.jbc.2024.105750 |