| First Author | Mukherjee J | Year | 2019 |
| Journal | Proc Natl Acad Sci U S A | Volume | 116 |
| Issue | 26 | Pages | 12863-12872 |
| PubMed ID | 31189591 | Mgi Jnum | J:276710 |
| Mgi Id | MGI:6316103 | Doi | 10.1073/pnas.1820737116 |
| Citation | Mukherjee J, et al. (2019) beta-Actin mRNA interactome mapping by proximity biotinylation. Proc Natl Acad Sci U S A 116(26):12863-12872 |
| abstractText | The molecular function and fate of mRNAs are controlled by RNA-binding proteins (RBPs). Identification of the interacting proteome of a specific mRNA in vivo remains very challenging, however. Based on the widely used technique of RNA tagging with MS2 aptamers for RNA visualization, we developed a RNA proximity biotinylation (RNA-BioID) technique by tethering biotin ligase (BirA*) via MS2 coat protein at the 3' UTR of endogenous MS2-tagged beta-actin mRNA in mouse embryonic fibroblasts. We demonstrate the dynamics of the beta-actin mRNA interactome by characterizing its changes on serum-induced localization of the mRNA. Apart from the previously known interactors, we identified more than 60 additional beta-actin-associated RBPs by RNA-BioID. Among these, the KH domain-containing protein FUBP3/MARTA2 has been shown to be required for beta-actin mRNA localization. We found that FUBP3 binds to the 3' UTR of beta-actin mRNA and is essential for beta-actin mRNA localization, but does not interact with the characterized beta-actin zipcode element. RNA-BioID provides a tool for identifying new mRNA interactors and studying the dynamic view of the interacting proteome of endogenous mRNAs in space and time. |
