Proximity labeling to detect RNA–protein interactions in live cells

Research output: Journal Publications and Reviews (RGC: 21, 22, 62)21_Publication in refereed journalNot applicablepeer-review

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Author(s)

Detail(s)

Original languageEnglish
Pages (from-to)1860-1868
Journal / PublicationFEBS Open Bio
Volume9
Issue number11
Online published27 Jul 2019
Publication statusPublished - Nov 2019

Abstract

RNA biology is orchestrated by the dynamic interactions of RNAs and RNA-binding proteins (RBPs). In the present study, we describe a new method of proximity-dependent protein labeling to detect RNA–protein interactions [RNA-bound protein proximity labeling (RBPL)]. We selected the well-studied RNA-binding protein PUF to examine the current proximity labeling enzymes birA* and APEX2. A new version of birA*, BASU, was used to validate that the PUF protein binds its RNA motif. We further optimized the RBPL labeling system using an inducible expression system. The RBPL (λN-BASU) labeling experiments exhibited high signal-to-noise ratios. We subsequently determined that RBPL (λN-BASU) is more suitable than RBPL (λN-APEX2) for the detection of RNA–protein interactions in live cells. Interestingly, our results also reveal that proximity labeling is probably capable of biotinylating proximate nascent peptide.

Research Area(s)

  • APEX2, birA*, nascent peptide, proximity labeling, RNA binding protein, RNA–protein interactions

Citation Format(s)

Proximity labeling to detect RNA–protein interactions in live cells. / Lu, Mingxing; Wei, Wencheng.

In: FEBS Open Bio, Vol. 9, No. 11, 11.2019, p. 1860-1868.

Research output: Journal Publications and Reviews (RGC: 21, 22, 62)21_Publication in refereed journalNot applicablepeer-review