A synergistic network of interactions promotes the formation of in vitro processing bodies and protects mRNA against decapping.

Abstract:

:Cellular liquid-liquid phase separation (LLPS) results in the formation of dynamic granules that play an important role in many biological processes. On a molecular level, the clustering of proteins into a confined space results from an indefinite network of intermolecular interactions. Here, we introduce and exploit a novel high-throughput bottom-up approach to study how the interactions between RNA, the Dcp1:Dcp2 mRNA decapping complex and the scaffolding proteins Edc3 and Pdc1 result in the formation of processing bodies. We find that the LLPS boundaries are close to physiological concentrations upon inclusion of multiple proteins and RNA. Within in vitro processing bodies the RNA is protected against endonucleolytic cleavage and the mRNA decapping activity is reduced, which argues for a role of processing bodies in temporary mRNA storage. Interestingly, the intrinsically disordered region (IDR) in the Edc3 protein emerges as a central hub for interactions with both RNA and mRNA decapping factors. In addition, the Edc3 IDR plays a role in the formation of irreversible protein aggregates that are potentially detrimental for cellular homeostasis. In summary, our data reveal insights into the mechanisms that lead to cellular LLPS and into the way this influences enzymatic activity.

journal_name

Nucleic Acids Res

journal_title

Nucleic acids research

authors

Schütz S,Nöldeke ER,Sprangers R

doi

10.1093/nar/gkx353

subject

Has Abstract

pub_date

2017-06-20 00:00:00

pages

6911-6922

issue

11

eissn

0305-1048

issn

1362-4962

pii

3787859

journal_volume

45

pub_type

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