Project Kartierung der Interaktome von lokalisierten mRNAs mittels Umgebung-Biotinylierung

Basic data

Title:
Kartierung der Interaktome von lokalisierten mRNAs mittels Umgebung-Biotinylierung
Duration:
01/05/2019 to 30/04/2022
Abstract / short description:
Function and fate of localized mRNAs are controlled by RNA binding proteins (RBPs) forming higher-order messenger ribonucleoprotein particles (mRNPs). Determining the proteome of a specific mRNP in vivo is still challenging. We have introduced a novel tool to study the interactome of mRNAs in vivo, RNA proximity biotinylation (RNA-BioID). RNA-BioID tethers a modified biotin ligase to an MS2-tagged RNA of choice that biotinylates proteins in the vicinity (about 20 nm diameter), and allows their affinity-based purification. We have demonstrated its practicality by mapping the dynamic proteome of the beta-actin mRNP in mouse embryonic fibroblasts (MEFs). We showed that this mRNP contains > 60 associated RBPs, including all six previously known interactors. In the second funding period, we will address three key questions of transport-competent mRNPs. Firstly, we will deepen the functional analysis of newly identified beta-actin interactors and their contribution to the local translation of beta-actin in MEFs. Secondly, we will characterize the interactome of beta-actin mRNA in differentiating neurons and compare it to the interactome in MEFs to map changes in RNA-associated proteins of a localizing mRNP. Thirdly, we aim to identify, characterize, and compare the interactome of an additional localized mRNA in MEFs (arp2) with that of beta-actin mRNA. Interactome mapping of mRNPs in cultured cells will be highly useful to study the dynamic interactions of proteins and mRNAs during their localization process.

Involved staff

Managers

Faculty of Science
University of Tübingen
Interfaculty Institute of Biochemistry (IFIB)
Interfaculty Institutes

Local organizational units

Interfaculty Institute of Biochemistry (IFIB)
Interfaculty Institutes
University of Tübingen

Funders

Bonn, Nordrhein-Westfalen, Germany
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