title: Rae1/YacP, a new endoribonuclease involved in ribosome-dependent mRNA decay inBacillus subtilis.
abstract: 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 ofBacillus subtilis, a founding member of the NYN (Nedd4-BP1/YacP nuclease) subfamily of PIN domain proteins, and identify potential substratesin vivoUnexpectedly, degradation of a characterised target mRNA was completely dependent on both its translation and reading frame. We provide evidence that Rae1 associates with theB. subtilisribosome and cleaves between specific codons of this mRNAin vivoCritically, we also demonstrate translation-dependent Rae1 cleavage of this substrate in a purified translation assayin vitroMultiple lines of evidence converge to suggest that Rae1 is an A-site endoribonuclease. We present a docking model of Rae1 bound to theB. subtilisribosomal 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.
authors.affiliation: UMR 8261 (CNRS - Univ. Paris Diderot, Sorbonne Paris Cité), Institut de Biologie Physico-Chimique, Paris, France.
authors.forename: Magali
authors.initials: M
authors.lastname: Leroy
authors.affiliation: UMR 8261 (CNRS - Univ. Paris Diderot, Sorbonne Paris Cité), Institut de Biologie Physico-Chimique, Paris, France.
authors.forename: Jérémie
authors.initials: J
authors.lastname: Piton
authors.affiliation: UMR 8261 (CNRS - Univ. Paris Diderot, Sorbonne Paris Cité), Institut de Biologie Physico-Chimique, Paris, France.
authors.forename: Laetitia
authors.initials: L
authors.lastname: Gilet
authors.affiliation: UMR 8261 (CNRS - Univ. Paris Diderot, Sorbonne Paris Cité), Institut de Biologie Physico-Chimique, Paris, France.
authors.forename: Olivier
authors.initials: O
authors.lastname: Pellegrini
authors.affiliation: Transcriptome and EpiGenome, Biomics Center for Innovation and Technological Research Institut Pasteur, Paris, France.
authors.forename: Caroline
authors.initials: C
authors.lastname: Proux
authors.affiliation: Transcriptome and EpiGenome, Biomics Center for Innovation and Technological Research Institut Pasteur, Paris, France.
authors.forename: Jean-Yves
authors.initials: JY
authors.lastname: Coppée
authors.affiliation: UMR 8261 (CNRS - Univ. Paris Diderot, Sorbonne Paris Cité), Institut de Biologie Physico-Chimique, Paris, France.
authors.forename: Sabine
authors.initials: S
authors.lastname: Figaro
authors.affiliation: UMR 8261 (CNRS - Univ. Paris Diderot, Sorbonne Paris Cité), Institut de Biologie Physico-Chimique, Paris, France condon@ibpc.fr.
authors.forename: Ciarán
authors.identifier: 0000-0002-2199-9621
authors.initials: C
authors.lastname: Condon
mesh_terms: D001412:Bacillus subtilis / Q000201:enzymology* / Q000378:metabolism*; D018360:Crystallography, X-Ray; D004722:Endoribonucleases / Q000737:chemistry* / Q000378:metabolism*; D008954:Models, Biological; D008958:Models, Molecular; D014176:Protein Biosynthesis; D011487:Protein Conformation; D020871:RNA Stability; D012270:Ribosomes / Q000378:metabolism*
chemical_list: D004722:Endoribonucleases
keywords: 
  Bacillus subtilis
  ; A‐site ribonuclease; NYN domain; RNA decay; translation