Open Access
Open access
Nucleic Acids Research, volume 45, issue 6, pages 3487-3502

Application of sorting and next generation sequencing to study 5'-UTR influence on translation efficiency in Escherichia coli

Evfratov Sergey A 1
Osterman Ilya A 2, 3
Pogorelskaya Alexandra M 1
Rubtsova Maria 2, 3
Semashko Tatiana 4
Kostryukova Elena S. 5
Mironov Andrey E. 2
Burnaev Evgeny 6
Krymova Ekaterina 7
GELFAND MICHAEL J. 2
Govorun Vadim M. 4
Bogdanov Alexey A. 2
Sergiev Petr V. 2, 3
Publication typeJournal Article
Publication date2016-11-28
Quartile SCImago
Q1
Quartile WOS
Q1
Impact factor14.9
ISSN03051048, 13624962
PubMed ID:  27899632
Genetics
Abstract
Abstract Yield of protein per translated mRNA may vary by four orders of magnitude. Many studies analyzed the influence of mRNA features on the translation yield. However, a detailed understanding of how mRNA sequence determines its propensity to be translated is still missing. Here, we constructed a set of reporter plasmid libraries encoding CER fluorescent protein preceded by randomized 5΄ untranslated regions (5΄-UTR) and Red fluorescent protein (RFP) used as an internal control. Each library was transformed into Escherchia coli cells, separated by efficiency of CER mRNA translation by a cell sorter and subjected to next generation sequencing. We tested efficiency of translation of the CER gene preceded by each of 48 natural 5΄-UTR sequences and introduced random and designed mutations into natural and artificially selected 5΄-UTRs. Several distinct properties could be ascribed to a group of 5΄-UTRs most efficient in translation. In addition to known ones, several previously unrecognized features that contribute to the translation enhancement were found, such as low proportion of cytidine residues, multiple SD sequences and AG repeats. The latter could be identified as translation enhancer, albeit less efficient than SD sequence in several natural 5΄-UTRs.

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Evfratov S. A. et al. Application of sorting and next generation sequencing to study 5'-UTR influence on translation efficiency in Escherichia coli // Nucleic Acids Research. 2016. Vol. 45. No. 6. pp. 3487-3502.
GOST all authors (up to 50) Copy
Evfratov S. A., Osterman I. A., Komarova E. S., Pogorelskaya A. M., Rubtsova M., Zatsepin T. S., Semashko T., Kostryukova E. S., Mironov A. E., Burnaev E., Krymova E., GELFAND M. J., Govorun V. M., Bogdanov A. A., Sergiev P. V., Dontsova O. A. Application of sorting and next generation sequencing to study 5'-UTR influence on translation efficiency in Escherichia coli // Nucleic Acids Research. 2016. Vol. 45. No. 6. pp. 3487-3502.
RIS |
Cite this
RIS Copy
TY - JOUR
DO - 10.1093/nar/gkw1141
UR - https://doi.org/10.1093%2Fnar%2Fgkw1141
TI - Application of sorting and next generation sequencing to study 5'-UTR influence on translation efficiency in Escherichia coli
T2 - Nucleic Acids Research
AU - Evfratov, Sergey A
AU - Pogorelskaya, Alexandra M
AU - Semashko, Tatiana
AU - Krymova, Ekaterina
AU - Dontsova, Olga A.
AU - Osterman, Ilya A
AU - Komarova, Ekaterina S
AU - Rubtsova, Maria
AU - Zatsepin, Timofei S.
AU - Kostryukova, Elena S.
AU - Mironov, Andrey E.
AU - Burnaev, Evgeny
AU - GELFAND, MICHAEL J.
AU - Govorun, Vadim M.
AU - Bogdanov, Alexey A.
AU - Sergiev, Petr V.
PY - 2016
DA - 2016/11/28 00:00:00
PB - Oxford University Press
SP - 3487-3502
IS - 6
VL - 45
PMID - 27899632
SN - 0305-1048
SN - 1362-4962
ER -
BibTex |
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BibTex Copy
@article{2016_Evfratov
author = {Sergey A Evfratov and Alexandra M Pogorelskaya and Tatiana Semashko and Ekaterina Krymova and Olga A. Dontsova and Ilya A Osterman and Ekaterina S Komarova and Maria Rubtsova and Timofei S. Zatsepin and Elena S. Kostryukova and Andrey E. Mironov and Evgeny Burnaev and MICHAEL J. GELFAND and Vadim M. Govorun and Alexey A. Bogdanov and Petr V. Sergiev},
title = {Application of sorting and next generation sequencing to study 5'-UTR influence on translation efficiency in Escherichia coli},
journal = {Nucleic Acids Research},
year = {2016},
volume = {45},
publisher = {Oxford University Press},
month = {nov},
url = {https://doi.org/10.1093%2Fnar%2Fgkw1141},
number = {6},
pages = {3487--3502},
doi = {10.1093/nar/gkw1141}
}
MLA
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MLA Copy
Evfratov, Sergey A., et al. “Application of sorting and next generation sequencing to study 5'-UTR influence on translation efficiency in Escherichia coli.” Nucleic Acids Research, vol. 45, no. 6, Nov. 2016, pp. 3487-3502. https://doi.org/10.1093%2Fnar%2Fgkw1141.
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