Evaluating ribosomal frameshifting in CCR5 mRNA decoding

PCR with reverse transcription (RT-PCR) of reporter RNAs showed that for cells transfected with the CCRS reporters, there was substantially less full-length cDNA and a simultaneous appearance of smaller molecular mass products (Fig. 1g). [...]the claim that the frameshift site is highly conserved am...

Full description

Saved in:
Bibliographic Details
Published inNature (London) Vol. 604; no. 7906; pp. E16 - E23
Main Authors Khan, Yousuf A., Loughran, Gary, Steckelberg, Anna-Lena, Brown, Katherine, Kiniry, Stephen J., Stewart, Hazel, Baranov, Pavel V., Kieft, Jeffrey S., Firth, Andrew E., Atkins, John F.
Format Journal Article
LanguageEnglish
Published London Nature Publishing Group UK 21.04.2022
Nature Publishing Group
Subjects
38/39
631/337/574/1789
631/45/500
Amino acid sequence
Amino acids
CCR5 protein
Complementary DNA
Efficiency
Frameshifting, Ribosomal - genetics
Gene expression
Humanities and Social Sciences
Laboratories
Matters Arising
multidisciplinary
Nucleic Acid Conformation
Polymerase chain reaction
Proteins
Reticulocytes
Reverse transcription
Ribosomes - genetics
Ribosomes - metabolism
RNA polymerase
RNA, Messenger - genetics
RNA, Messenger - metabolism
Science
Science (multidisciplinary)
Online AccessGet full text

Cover

More Information
Summary:PCR with reverse transcription (RT-PCR) of reporter RNAs showed that for cells transfected with the CCRS reporters, there was substantially less full-length cDNA and a simultaneous appearance of smaller molecular mass products (Fig. 1g). [...]the claim that the frameshift site is highly conserved among higher primates does not indicate that the (putative) frameshift signal is subject to purifying selection. [...]a rabbit reticulocyte lysate in vitro translation system was programmed with capped RNAs (Fig. 2j). Methods Alignment of CCR5 slippery sites The NCBI RefSeq database was searched with tblastn using the human CCR5 amino acid sequence derived from mRNA accession NM_000579.3 as query on 4 December 2018 using default parameters, with the number of hits expanded from 100 to 500.
Bibliography:ObjectType-Article-1
SourceType-Scholarly Journals-1
ObjectType-Feature-2
content type line 14
ObjectType-Article-2
ObjectType-Correspondence-1
content type line 23
Author contribution statement: The splicing work, which was initiated in 2016, was performed by A-L.S and J.S.K.; two teams, one involving G.L. and J.F.A, and the other involving Y.A.K., K.B., H.S. and A.E.F. semi-independently performed the frameshifting analysis; S.J.K. and P.V.B., with input from A.E.F. performed the ribosome profiling analysis. After initial independent work of the teams involved, Y.A.K. and J.F.A. coordinated their combination. Y.A.K. and G.L. had major roles in preparing the manuscript with contributions and refinements from all other authors.
ISSN:0028-0836
1476-4687
1476-4687
DOI:10.1038/s41586-022-04627-y