Biochemical approach for isolation of polyadenylated RNAs with bound proteins from yeast.
STAR Protoc
; 2(4): 100929, 2021 12 17.
Article
en En
| MEDLINE
| ID: mdl-34766032
ABSTRACT
In vivo characterization of RNA-protein interactions is the key for understanding RNA regulatory mechanisms. Herein, we describe a protocol for detection of proteins interacting with polyadenylated RNAs in the yeast Saccharomyces cerevisiae. Proteins are crosslinked to nucleic acids in vivo by ultraviolet (UV) irradiation of cells, and poly(A)-containing RNAs with bound proteins are isolated from cell lysates using oligo[dT]25 beads. RBPs can be detected by immunoblot analysis or with mass spectrometry to define the mRNA-binding proteome (mRBPome) and its changes under stress. For complete details on the use and execution of this protocol, please refer to Matia-González et al. (2021, 2015).
Palabras clave
Texto completo:
1
Colección:
01-internacional
Base de datos:
MEDLINE
Asunto principal:
Espectrometría de Masas
/
ARN de Hongos
/
ARN Mensajero
/
Proteínas de Unión al ARN
/
Proteínas de Saccharomyces cerevisiae
Idioma:
En
Revista:
STAR Protoc
Año:
2021
Tipo del documento:
Article
País de afiliación:
Reino Unido