title: Human Cells Require Non-stop Ribosome Rescue Activity in Mitochondria.
abstract: Bacteria use trans-translation and the alternative rescue factors ArfA (P36675) and ArfB (Q9A8Y3) to hydrolyze peptidyl-tRNA on ribosomes that stall near the 3' end of an mRNA during protein synthesis. The eukaryotic protein ICT1 (Q14197) is homologous to ArfB. In vitro ribosome rescue assays of human ICT1 and Caulobacter crescentus ArfB showed that these proteins have the same activity and substrate specificity. Both ArfB and ICT1 hydrolyze peptidyl-tRNA on nonstop ribosomes or ribosomes stalled with ≤6 nucleotides extending past the A site, but are unable to hydrolyze peptidyl-tRNA when the mRNA extends ≥14 nucleotides past the A site. ICT1 provided sufficient ribosome rescue activity to support viability in C. crescentus cells that lacked both trans-translation and ArfB. Likewise, expression of ArfB protected human cells from death when ICT1 was silenced with siRNA. These data indicate that ArfB and ICT1 are functionally interchangeable, and demonstrate that ICT1 is a ribosome rescue factor. Because ICT1 is essential in human cells, these results suggest that ribosome rescue activity in mitochondria is required in humans.
authors.affiliation: Department of Biochemistry and Molecular Biology, The Pennsylvania State University, University Park, Pennsylvania, United States of America.
authors.forename: Heather A
authors.initials: HA
authors.lastname: Feaga
authors.affiliation: Department of Veterinary and Biomedical Science, The Pennsylvania State University, University Park, Pennsylvania, United States of America.
authors.forename: Michael D
authors.initials: MD
authors.lastname: Quickel
authors.affiliation: Department of Veterinary and Biomedical Science, The Pennsylvania State University, University Park, Pennsylvania, United States of America.
authors.forename: Pamela A
authors.initials: PA
authors.lastname: Hankey-Giblin
authors.affiliation: Department of Biochemistry and Molecular Biology, The Pennsylvania State University, University Park, Pennsylvania, United States of America.
authors.forename: Kenneth C
authors.initials: KC
authors.lastname: Keiler
mesh_terms: D016935:Caulobacter crescentus / Q000235:genetics; D057809:HEK293 Cells; D006801:Humans; D008928:Mitochondria / Q000235:genetics* / Q000378:metabolism; D014176:Protein Biosynthesis / Q000235:genetics*; D011506:Proteins / Q000235:genetics* / Q000378:metabolism; D012333:RNA, Messenger / Q000235:genetics; D012346:RNA, Transfer, Amino Acyl / Q000235:genetics; D016601:RNA-Binding Proteins / Q000235:genetics; D012269:Ribosomal Proteins; D012270:Ribosomes / Q000235:genetics* / Q000378:metabolism; D014994:Xylose / Q000378:metabolism
chemical_list: C098021:MRPL58 protein, human; D011506:Proteins; D012333:RNA, Messenger; D012346:RNA, Transfer, Amino Acyl; D016601:RNA-Binding Proteins; D012269:Ribosomal Proteins; C019037:tRNA, peptidyl-; D014994:Xylose