Insights into RNA biology from an atlas of mammalian mRNA-binding proteins.
RNA-binding proteins (RBPs) determine RNA fate from synthesis to decay. Employing two complementary protocols for covalent UV crosslinking of RBPs to RNA, we describe a systematic, unbiased, and comprehensive approach, termed "interactome capture," to define the mRNA interactome of proliferating human HeLa cells. We identify 860 proteins that qualify as RBPs by biochemical and statistical criteria, adding more than 300 RBPs to those previously known and shedding light on RBPs in disease, RNA-binding enzymes of intermediary metabolism, RNA-binding kinases, and RNA-binding architectures. Unexpectedly, we find that many proteins of the HeLa mRNA interactome are highly intrinsically disordered and enriched in short repetitive amino acid motifs. Interactome capture is broadly applicable to study mRNA interactome composition and dynamics in varied biological settings.
European Molecular Biology Laboratory, Heidelberg, Germany.
Alfredo
A
Castello
Bernd
B
Fischer
Katrin
K
Eichelbaum
Rastislav
R
Horos
Benedikt M
BM
Beckmann
Claudia
C
Strein
Norman E
NE
Davey
David T
DT
Humphreys
Thomas
T
Preiss
Lars M
LM
Steinmetz
Jeroen
J
Krijgsveld
Matthias W
MW
Hentze
D000818:Animals; D006367:HeLa Cells; D006801:Humans; D040901:Proteomics / Q000379:methods*; D012333:RNA, Messenger / Q000378:metabolism*; D016601:RNA-Binding Proteins / Q000302:isolation & purification* / Q000378:metabolism
D012333:RNA, Messenger; D016601:RNA-Binding Proteins