Proximity labeling to detect RNA-protein interactions in live cells

Lu, Mingxing1; Wei, Wencheng2

2019-09-25

10.1002/2211-5463.12706

FEBS Open Bio   影响因子和分区

2211-5463

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 (lambda N-BASU) labeling experiments exhibited high signal-to-noise ratios. We subsequently determined that RBPL (lambda N-BASU) is more suitable than RBPL (lambda N-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.

APEX2 birA* nascent peptide proximity labeling RNA binding protein RNA-protein interactions

SCI

英语

其他

Biochemistry & Molecular Biology

Biochemistry & Molecular Biology

WOS:000490261000001

WILEY

Web of Science

1.City Univ Hong Kong, Coll Vet Med & Life Sci, Dept Biomed Sci, Kowloon Tong, 83 Tat Chee Ave, Hong Kong 999077, Peoples R China
2.Southern Univ Sci & Technol, Dept Biol, Shenzhen, Peoples R China

Lu, Mingxing,Wei, Wencheng. Proximity labeling to detect RNA-protein interactions in live cells[J]. FEBS Open Bio,2019,9(11):1860-1868.

Lu, Mingxing,&Wei, Wencheng.(2019).Proximity labeling to detect RNA-protein interactions in live cells.FEBS Open Bio,9(11),1860-1868.

Lu, Mingxing,et al."Proximity labeling to detect RNA-protein interactions in live cells".FEBS Open Bio 9.11(2019):1860-1868.