Insertion mutagenesis to increase secondary structure within the 5' noncoding region of a eukaryotic mRNA reduces translational efficiency - PubMed (original) (raw)
Insertion mutagenesis to increase secondary structure within the 5' noncoding region of a eukaryotic mRNA reduces translational efficiency
J Pelletier et al. Cell. 1985 Mar.
Abstract
The thymidine kinase gene of herpes simplex virus 1 was mutated by inserting oligodeoxynucleotide linkers into the region of the gene corresponding to the 5' untranslated portion of the mRNA. These linkers, when transcribed into mRNA, might be expected to form hairpin loops and hence to increase the secondary structure of the 5' end of the mRNA. Thymidine kinase insertion derivatives were examined in vivo and in vitro to determine translational efficiency. For the in vivo studies, thymidine kinase insertion derivatives were transfected into thymidine kinase deficient L cells alone and together with a selectable dominant marker, or were assayed in the COS-1 transient expression system. For in vitro studies, thymidine kinase insertion derivatives were subcloned into pSP64. Capped transcripts were analyzed for their ability to bind ribosomes and translate in rabbit reticulocyte lysates and wheat-germ extracts. The results demonstrate that translation efficiency is decreased as the number of linkers is increased and support the view that excessive secondary structure at the 5' end of eukaryotic mRNA impedes translation.
Similar articles
- Expression of herpes simplex virus type I thymidine kinase gene in Escherichia coli.
Garapin AC, Colbère-Garapin F, Cohen-Solal M, Horodniceanu F, Kourilsky P. Garapin AC, et al. Proc Natl Acad Sci U S A. 1981 Feb;78(2):815-9. doi: 10.1073/pnas.78.2.815. Proc Natl Acad Sci U S A. 1981. PMID: 6262774 Free PMC article. - Nucleotide sequence of the thymidine kinase gene of herpes simplex virus type 1.
Wagner MJ, Sharp JA, Summers WC. Wagner MJ, et al. Proc Natl Acad Sci U S A. 1981 Mar;78(3):1441-5. doi: 10.1073/pnas.78.3.1441. Proc Natl Acad Sci U S A. 1981. PMID: 6262799 Free PMC article. - Characterization of eukaryotic transcriptional control signals by assay of herpes simplex virus type 1 thymidine kinase.
Lang JC, Wilkie NM, Spandidos DA. Lang JC, et al. J Gen Virol. 1983 Dec;64 ( Pt 12):2679-96. doi: 10.1099/0022-1317-64-12-2679. J Gen Virol. 1983. PMID: 6319553 - Control of expression of the herpes simplex virus thymidine kinase gene in biochemically transformed cells.
El Kareh A, Silverstein S, Smiley J. El Kareh A, et al. J Gen Virol. 1984 Jan;65 ( Pt 1):19-36. doi: 10.1099/0022-1317-65-1-19. J Gen Virol. 1984. PMID: 6319567 - Construction of a dominant selective marker useful for gene transfer studies in animal cells.
Colbère-Garapin F, Horodniceanu F, Kourilsky P, Garapin AC. Colbère-Garapin F, et al. Dev Biol Stand. 1981;50:323-6. Dev Biol Stand. 1981. PMID: 6281111
Cited by
- From structural design to delivery: mRNA therapeutics for cancer immunotherapy.
Zhou F, Huang L, Li S, Yang W, Chen F, Cai Z, Liu X, Xu W, Lehto VP, Lächelt U, Huang R, Shi Y, Lammers T, Tao W, Xu ZP, Wagner E, Xu Z, Yu H. Zhou F, et al. Exploration (Beijing). 2023 Nov 17;4(2):20210146. doi: 10.1002/EXP.20210146. eCollection 2024 Apr. Exploration (Beijing). 2023. PMID: 38855617 Free PMC article. Review. - Differences in 5'untranslated regions highlight the importance of translational regulation of dosage sensitive genes.
Wieder N, D'Souza EN, Martin-Geary AC, Lassen FH, Talbot-Martin J, Fernandes M, Chothani SP, Rackham OJL, Schafer S, Aspden JL, MacArthur DG, Davies RW, Whiffin N. Wieder N, et al. Genome Biol. 2024 Apr 29;25(1):111. doi: 10.1186/s13059-024-03248-0. Genome Biol. 2024. PMID: 38685090 Free PMC article. - Host-like RNA Elements Regulate Virus Translation.
Khan D, Fox PL. Khan D, et al. Viruses. 2024 Mar 20;16(3):468. doi: 10.3390/v16030468. Viruses. 2024. PMID: 38543832 Free PMC article. Review. - Combinatorial optimization of gene expression through recombinase-mediated promoter and terminator shuffling in yeast.
Cautereels C, Smets J, Bircham P, De Ruysscher D, Zimmermann A, De Rijk P, Steensels J, Gorkovskiy A, Masschelein J, Verstrepen KJ. Cautereels C, et al. Nat Commun. 2024 Feb 7;15(1):1112. doi: 10.1038/s41467-024-44997-7. Nat Commun. 2024. PMID: 38326309 Free PMC article. - The regulatory landscape of 5' UTRs in translational control during zebrafish embryogenesis.
Reimão-Pinto MM, Castillo-Hair SM, Seelig G, Schier AF. Reimão-Pinto MM, et al. bioRxiv [Preprint]. 2023 Nov 23:2023.11.23.568470. doi: 10.1101/2023.11.23.568470. bioRxiv. 2023. PMID: 38045294 Free PMC article. Preprint.
Publication types
MeSH terms
Substances
LinkOut - more resources
Full Text Sources
Other Literature Sources