Research output: Contribution to journal › Article › peer-review
Structural rearrangements in mRNA upon its binding to human 80S ribosomes revealed by EPR spectroscopy. / Malygin, Alexey A.; Graifer, Dmitri M.; Meschaninova, Maria I. et al.
In: Nucleic Acids Research, Vol. 46, No. 2, 25.01.2018, p. 897-904.Research output: Contribution to journal › Article › peer-review
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TY - JOUR
T1 - Structural rearrangements in mRNA upon its binding to human 80S ribosomes revealed by EPR spectroscopy
AU - Malygin, Alexey A.
AU - Graifer, Dmitri M.
AU - Meschaninova, Maria I.
AU - Venyaminova, Alya G.
AU - Timofeev, Ivan O.
AU - Kuzhelev, Andrey A.
AU - Krumkacheva, Olesya A.
AU - Fedin, Matvey V.
AU - Karpova, Galina G.
AU - Bagryanskaya, Elena G.
N1 - Publisher Copyright: © The Author(s) 2017.
PY - 2018/1/25
Y1 - 2018/1/25
N2 - The model mRNA (MR), 11-mer RNA containing two nitroxide spin labels at the 5'- and 3 -terminal nucleotides and prone to form a stable homodimer (MR)2, was used for Electron Paramagnetic Resonance study of structural rearrangements in mRNA occurring upon its binding to human 80S ribosomes. The formation of two different types of ribosomal complexes with MR was observed. First, there were stable complexes where MR was fixed in the ribosomal mRNA-binding channel by the codon-anticodon interaction(s) with cognate tRNA(s). Second, we for the first time detected complexes assembled without tRNA due to the binding of MR most likely to an exposed peptide of ribosomal protein uS3 away from the mRNA channel. The analysis of interspin distances allowed the conclusion that 80S ribosomes facilitate dissociation of the duplex (MR)2: the equilibrium between the duplex and the single-stranded MR shifts to MR due to its efficient binding with ribosomes. Furthermore, we observed a significant influence of tRNA bound at the ribosomal exit (E) and/or aminoacyl (A) sites on the stability of ribosomal complexes. Our findings showed that a part of mRNA bound in the ribosome channel, which is not involved in codon-anticodon interactions, has more degrees of freedom than that interacting with tRNAs.
AB - The model mRNA (MR), 11-mer RNA containing two nitroxide spin labels at the 5'- and 3 -terminal nucleotides and prone to form a stable homodimer (MR)2, was used for Electron Paramagnetic Resonance study of structural rearrangements in mRNA occurring upon its binding to human 80S ribosomes. The formation of two different types of ribosomal complexes with MR was observed. First, there were stable complexes where MR was fixed in the ribosomal mRNA-binding channel by the codon-anticodon interaction(s) with cognate tRNA(s). Second, we for the first time detected complexes assembled without tRNA due to the binding of MR most likely to an exposed peptide of ribosomal protein uS3 away from the mRNA channel. The analysis of interspin distances allowed the conclusion that 80S ribosomes facilitate dissociation of the duplex (MR)2: the equilibrium between the duplex and the single-stranded MR shifts to MR due to its efficient binding with ribosomes. Furthermore, we observed a significant influence of tRNA bound at the ribosomal exit (E) and/or aminoacyl (A) sites on the stability of ribosomal complexes. Our findings showed that a part of mRNA bound in the ribosome channel, which is not involved in codon-anticodon interactions, has more degrees of freedom than that interacting with tRNAs.
KW - Anticodon/metabolism
KW - Binding Sites
KW - Codon/metabolism
KW - Electron Spin Resonance Spectroscopy/methods
KW - Humans
KW - Nucleic Acid Conformation
KW - Protein Binding
KW - RNA, Messenger/chemistry
KW - RNA, Transfer/chemistry
KW - Ribosomal Proteins/metabolism
KW - Ribosomes/metabolism
KW - Spin Labels
UR - http://www.scopus.com/inward/record.url?scp=85044640684&partnerID=8YFLogxK
U2 - 10.1093/nar/gkx1136
DO - 10.1093/nar/gkx1136
M3 - Article
C2 - 29156000
AN - SCOPUS:85044640684
VL - 46
SP - 897
EP - 904
JO - Nucleic Acids Research
JF - Nucleic Acids Research
SN - 0305-1048
IS - 2
ER -
ID: 12282750