Functional identification of microRNA-centered complexes in C. elegans
Abstract microRNAs (miRNAs) are crucial for normal development and physiology. To identify factors that might coordinate with miRNAs to regulate gene expression, we used 2′O-methylated oligonucleotides to precipitate Caenorhabditis elegans let-7, miR-58, and miR-2 miRNAs and the associated proteins....
| Main Authors: | , , , , , |
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| Format: | Article |
| Language: | English |
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Nature Portfolio
2022-05-01
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| Series: | Scientific Reports |
| Online Access: | https://doi.org/10.1038/s41598-022-10771-2 |
| _version_ | 1818195662998601728 |
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| author | Shilpa Hebbar Ganesh Panzade Ajay A. Vashisht James A. Wohlschlegel Isana Veksler-Lublinsky Anna Y. Zinovyeva |
| author_facet | Shilpa Hebbar Ganesh Panzade Ajay A. Vashisht James A. Wohlschlegel Isana Veksler-Lublinsky Anna Y. Zinovyeva |
| author_sort | Shilpa Hebbar |
| collection | DOAJ |
| description | Abstract microRNAs (miRNAs) are crucial for normal development and physiology. To identify factors that might coordinate with miRNAs to regulate gene expression, we used 2′O-methylated oligonucleotides to precipitate Caenorhabditis elegans let-7, miR-58, and miR-2 miRNAs and the associated proteins. A total of 211 proteins were identified through mass-spectrometry analysis of miRNA co-precipitates, which included previously identified interactors of key miRNA pathway components. Gene ontology analysis of the identified interactors revealed an enrichment for RNA binding proteins, suggesting that we captured proteins that may be involved in mRNA lifecycle. To determine which miRNA interactors are important for miRNA activity, we used RNAi to deplete putative miRNA co-factors in animals with compromised miRNA activity and looked for alterations of the miRNA mutant phenotypes. Depletion of 25 of 39 tested genes modified the miRNA mutant phenotypes in three sensitized backgrounds. Modulators of miRNA phenotypes ranged from RNA binding proteins RBD-1 and CEY-1 to metabolic factors such as DLST-1 and ECH-5, among others. The observed functional interactions suggest widespread coordination of these proteins with miRNAs to ultimately regulate gene expression. This study provides a foundation for future investigations aimed at deciphering the molecular mechanisms of miRNA-mediated gene regulation. |
| first_indexed | 2024-12-12T01:21:45Z |
| format | Article |
| id | doaj.art-0d21b2655acd4ddb94d7327df8708ac2 |
| institution | Directory Open Access Journal |
| issn | 2045-2322 |
| language | English |
| last_indexed | 2024-12-12T01:21:45Z |
| publishDate | 2022-05-01 |
| publisher | Nature Portfolio |
| record_format | Article |
| series | Scientific Reports |
| spelling | doaj.art-0d21b2655acd4ddb94d7327df8708ac22022-12-22T00:43:12ZengNature PortfolioScientific Reports2045-23222022-05-0112111510.1038/s41598-022-10771-2Functional identification of microRNA-centered complexes in C. elegansShilpa Hebbar0Ganesh Panzade1Ajay A. Vashisht2James A. Wohlschlegel3Isana Veksler-Lublinsky4Anna Y. Zinovyeva5Division of Biology, Kansas State UniversityDivision of Biology, Kansas State UniversityDepartment of Biological Chemistry, David Geffen School of Medicine, University of CaliforniaDepartment of Biological Chemistry, David Geffen School of Medicine, University of CaliforniaDepartment of Software and Information Systems Engineering, Ben-Gurion University of the NegevDivision of Biology, Kansas State UniversityAbstract microRNAs (miRNAs) are crucial for normal development and physiology. To identify factors that might coordinate with miRNAs to regulate gene expression, we used 2′O-methylated oligonucleotides to precipitate Caenorhabditis elegans let-7, miR-58, and miR-2 miRNAs and the associated proteins. A total of 211 proteins were identified through mass-spectrometry analysis of miRNA co-precipitates, which included previously identified interactors of key miRNA pathway components. Gene ontology analysis of the identified interactors revealed an enrichment for RNA binding proteins, suggesting that we captured proteins that may be involved in mRNA lifecycle. To determine which miRNA interactors are important for miRNA activity, we used RNAi to deplete putative miRNA co-factors in animals with compromised miRNA activity and looked for alterations of the miRNA mutant phenotypes. Depletion of 25 of 39 tested genes modified the miRNA mutant phenotypes in three sensitized backgrounds. Modulators of miRNA phenotypes ranged from RNA binding proteins RBD-1 and CEY-1 to metabolic factors such as DLST-1 and ECH-5, among others. The observed functional interactions suggest widespread coordination of these proteins with miRNAs to ultimately regulate gene expression. This study provides a foundation for future investigations aimed at deciphering the molecular mechanisms of miRNA-mediated gene regulation.https://doi.org/10.1038/s41598-022-10771-2 |
| spellingShingle | Shilpa Hebbar Ganesh Panzade Ajay A. Vashisht James A. Wohlschlegel Isana Veksler-Lublinsky Anna Y. Zinovyeva Functional identification of microRNA-centered complexes in C. elegans Scientific Reports |
| title | Functional identification of microRNA-centered complexes in C. elegans |
| title_full | Functional identification of microRNA-centered complexes in C. elegans |
| title_fullStr | Functional identification of microRNA-centered complexes in C. elegans |
| title_full_unstemmed | Functional identification of microRNA-centered complexes in C. elegans |
| title_short | Functional identification of microRNA-centered complexes in C. elegans |
| title_sort | functional identification of microrna centered complexes in c elegans |
| url | https://doi.org/10.1038/s41598-022-10771-2 |
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