Dataset Information

β-actin RNA-BioID

ABSTRACT: Function and fate of mRNAs is controlled by RNA binding proteins (RBPs) but determining the proteome of a specific mRNA in vivo is still challenging. RNA proximity biotinylation on the transported β-actin mRNA tagged with MS2 aptamers (RNA-BioID) is used to characterize the dynamic proteome of the β-actin mRNP in mouse embryonic fibroblasts (MEFs). We have identified > 60 β-actin associated RBPs including all six previously known as well as novel interactors. By investigating the dynamics of the β-actin mRNP in MEFs, we expand the set of β-actin mRNA associated RBPs and characterize the changes of the interacting proteome upon serum-induced mRNA localization. We report that the KH-domain containing protein FUBP3 represents a new β-actin associated RBP that binds to its 3’-untranslated region outside the known RNA localization element but is required for β-actin RNA localization. RNA-BioID will allow to obtain a dynamic view on the composition of endogenous mRNPs.

INSTRUMENT(S): LTQ Orbitrap Elite

ORGANISM(S): Mus Musculus (mouse)

TISSUE(S): Embryonic Fibroblast

SUBMITTER: Mirita Franz-Wachtel

LAB HEAD: Ralf-Peter Jansen

PROVIDER: PXD010694 | Pride | 2020-03-12

REPOSITORIES: Pride

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Dataset's files

Source:

			Action	DRS
	20180315_CO_0651RaJa_R57.raw	Raw
	20180315_CO_0651RaJa_R58.raw	Raw
	20180315_CO_0651RaJa_R59.raw	Raw
	20180315_CO_0651RaJa_R63.raw	Raw
	20180315_CO_0651RaJa_R64.raw	Raw

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Publications

β-Actin mRNA interactome mapping by proximity biotinylation.

Mukherjee Joyita J Hermesh Orit O Eliscovich Carolina C Nalpas Nicolas N Franz-Wachtel Mirita M Maček Boris B Jansen Ralf-Peter RP

Proceedings of the National Academy of Sciences of the United States of America 20190612 26

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 β-actin mRNA in mouse embryonic fibroblasts. We demonstr ...[more]

PMID: 31189591

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