title: N(6)-methyladenosine in mRNA disrupts tRNA selection and translation-elongation dynamics.
abstract: N(6)-methylation of adenosine (forming m(6)A) is the most abundant post-transcriptional modification within the coding region of mRNA, but its role during translation remains unknown. Here, we used bulk kinetic and single-molecule methods to probe the effect of m(6)A in mRNA decoding. Although m(6)A base-pairs with uridine during decoding, as shown by X-ray crystallographic analyses of Thermus thermophilus ribosomal complexes, our measurements in an Escherichia coli translation system revealed that m(6)A modification of mRNA acts as a barrier to tRNA accommodation and translation elongation. The interaction between an m(6)A-modified codon and cognate tRNA echoes the interaction between a near-cognate codon and tRNA, because delay in tRNA accommodation depends on the position and context of m(6)A within codons and on the accuracy level of translation. Overall, our results demonstrate that chemical modification of mRNA can change translational dynamics.
authors.affiliation: Department of Structural Biology, Stanford University School of Medicine, Stanford, California, USA.|Department of Applied Physics, Stanford University, Stanford, California, USA.
authors.forename: Junhong
authors.identifier: http://orcid.org/0000-0001-9291-5977
authors.initials: J
authors.lastname: Choi
authors.affiliation: Department of Cell and Molecular Biology, Biomedical Center, Uppsala University, Uppsala, Sweden.
authors.forename: Ka-Weng
authors.initials: KW
authors.lastname: Ieong
authors.affiliation: Stanford PULSE Institute, SLAC National Accelerator Laboratory, Menlo Park, California, USA.|Stanford Synchrotron Radiation Lightsource, SLAC National Accelerator Laboratory, Menlo Park, California, USA.
authors.forename: Hasan
authors.identifier: http://orcid.org/0000-0002-9135-5397
authors.initials: H
authors.lastname: Demirci
authors.affiliation: Department of Structural Biology, Stanford University School of Medicine, Stanford, California, USA.|Department of Applied Physics, Stanford University, Stanford, California, USA.
authors.forename: Jin
authors.initials: J
authors.lastname: Chen
authors.affiliation: Department of Structural Biology, Stanford University School of Medicine, Stanford, California, USA.
authors.forename: Alexey
authors.initials: A
authors.lastname: Petrov
authors.affiliation: Department of Structural Biology, Stanford University School of Medicine, Stanford, California, USA.|Program in Biophysics, Stanford University, Stanford, California, USA.
authors.forename: Arjun
authors.initials: A
authors.lastname: Prabhakar
authors.affiliation: Department of Structural Biology, Stanford University School of Medicine, Stanford, California, USA.
authors.forename: Seán E
authors.initials: SE
authors.lastname: O'Leary
authors.affiliation: Cancer Research Center, Chaim Sheba Medical Center, Tel Hashomer, Israel.
authors.forename: Dan
authors.initials: D
authors.lastname: Dominissini
authors.affiliation: Cancer Research Center, Chaim Sheba Medical Center, Tel Hashomer, Israel.|Israel &Sackler School of Medicine, Tel Aviv University, Tel Aviv, Israel.
authors.forename: Gideon
authors.initials: G
authors.lastname: Rechavi
authors.affiliation: Stanford Synchrotron Radiation Lightsource, SLAC National Accelerator Laboratory, Menlo Park, California, USA.
authors.forename: S Michael
authors.initials: SM
authors.lastname: Soltis
authors.affiliation: Department of Cell and Molecular Biology, Biomedical Center, Uppsala University, Uppsala, Sweden.
authors.forename: Måns
authors.initials: M
authors.lastname: Ehrenberg
authors.affiliation: Department of Structural Biology, Stanford University School of Medicine, Stanford, California, USA.
authors.forename: Joseph D
authors.initials: JD
authors.lastname: Puglisi
mesh_terms: D000241:Adenosine / Q000031:analogs & derivatives* / Q000032:analysis / Q000235:genetics; D003062:Codon; D018360:Crystallography, X-Ray; D004926:Escherichia coli / Q000737:chemistry / Q000235:genetics*; D014176:Protein Biosynthesis; D012329:RNA, Bacterial / Q000737:chemistry / Q000235:genetics*; D012333:RNA, Messenger / Q000737:chemistry / Q000235:genetics*; D012343:RNA, Transfer / Q000737:chemistry / Q000235:genetics*; D016963:Thermus thermophilus / Q000737:chemistry / Q000235:genetics*
chemical_list: D003062:Codon; D012329:RNA, Bacterial; D012333:RNA, Messenger; D012343:RNA, Transfer; C010223:N-methyladenosine; D000241:Adenosine