Premium
Rae1/YacP, a new endoribonuclease involved in ribosome‐dependent mRNA decay in Bacillus subtilis
Author(s) -
Leroy Magali,
Piton Jérémie,
Gilet Laetitia,
Pellegrini Olivier,
Proux Caroline,
Coppée JeanYves,
Figaro Sabine,
Condon Ciarán
Publication year - 2017
Publication title -
the embo journal
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 7.484
H-Index - 392
eISSN - 1460-2075
pISSN - 0261-4189
DOI - 10.15252/embj.201796540
Subject(s) - biology , endoribonuclease , microbiology and biotechnology , bacillus subtilis , start codon , messenger rna , rna , ribosome , rnase p , ribosomal binding site , genetics , gene , bacteria
The PIN domain plays a central role in cellular RNA biology and is involved in processes as diverse as rRNA maturation, mRNA decay and telomerase function. Here, we solve the crystal structure of the Rae1 (YacP) protein of Bacillus subtilis , a founding member of the NYN (Nedd4‐BP1/YacP nuclease) subfamily of PIN domain proteins, and identify potential substrates in vivo . Unexpectedly, degradation of a characterised target mRNA was completely dependent on both its translation and reading frame. We provide evidence that Rae1 associates with the B. subtilis ribosome and cleaves between specific codons of this mRNA in vivo . Critically, we also demonstrate translation‐dependent Rae1 cleavage of this substrate in a purified translation assay in vitro . Multiple lines of evidence converge to suggest that Rae1 is an A‐site endoribonuclease. We present a docking model of Rae1 bound to the B. subtilis ribosomal A‐site that is consistent with this hypothesis and show that Rae1 cleaves optimally immediately upstream of a lysine codon (AAA or AAG) in vivo .