mRNA stem-loops can pause the ribosome by hindering A-site tRNA binding

Although the elongating ribosome is an efficient helicase, certain mRNA stem-loop structures are known to impede ribosome movement along mRNA and stimulate programmed ribosome frameshifting via mechanisms that are not well understood. Using biochemical and single-molecule Förster resonance energy tr...
Ausführliche Beschreibung

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Autor*in:	Chen Bao [verfasserIn] Sarah Loerch [verfasserIn] Clarence Ling [verfasserIn] Andrei A Korostelev [verfasserIn] Nikolaus Grigorieff [verfasserIn] Dmitri N Ermolenko [verfasserIn]

Format:	E-Artikel
Sprache:	Englisch

Erschienen:	2020

Schlagwörter:	ribosome translation mRNA smFRET cryo-EM

Übergeordnetes Werk:	In: eLife - eLife Sciences Publications Ltd, 2013, 9(2020)
Übergeordnetes Werk:	volume:9 ; year:2020

Links:	Link aufrufen Link aufrufen Link aufrufen Journal toc

DOI / URN:	10.7554/eLife.55799

Katalog-ID:	DOAJ004384385

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520			\|a Although the elongating ribosome is an efficient helicase, certain mRNA stem-loop structures are known to impede ribosome movement along mRNA and stimulate programmed ribosome frameshifting via mechanisms that are not well understood. Using biochemical and single-molecule Förster resonance energy transfer (smFRET) experiments, we studied how frameshift-inducing stem-loops from E. coli dnaX mRNA and the gag-pol transcript of Human Immunodeficiency Virus (HIV) perturb translation elongation. We find that upon encountering the ribosome, the stem-loops strongly inhibit A-site tRNA binding and ribosome intersubunit rotation that accompanies translation elongation. Electron cryo-microscopy (cryo-EM) reveals that the HIV stem-loop docks into the A site of the ribosome. Our results suggest that mRNA stem-loops can transiently escape the ribosome helicase by binding to the A site. Thus, the stem-loops can modulate gene expression by sterically hindering tRNA binding and inhibiting translation elongation.
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Indexfelder

author_variant	c b cb s l sl c l cl a a k aak n g ng d n e dne
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allfields	10.7554/eLife.55799 doi (DE-627)DOAJ004384385 (DE-599)DOAJ716549eb2dca4eb8a1914d4e9a7819df DE-627 ger DE-627 rakwb eng QH301-705.5 Chen Bao verfasserin aut mRNA stem-loops can pause the ribosome by hindering A-site tRNA binding 2020 Text txt rdacontent Computermedien c rdamedia Online-Ressource cr rdacarrier Although the elongating ribosome is an efficient helicase, certain mRNA stem-loop structures are known to impede ribosome movement along mRNA and stimulate programmed ribosome frameshifting via mechanisms that are not well understood. Using biochemical and single-molecule Förster resonance energy transfer (smFRET) experiments, we studied how frameshift-inducing stem-loops from E. coli dnaX mRNA and the gag-pol transcript of Human Immunodeficiency Virus (HIV) perturb translation elongation. We find that upon encountering the ribosome, the stem-loops strongly inhibit A-site tRNA binding and ribosome intersubunit rotation that accompanies translation elongation. Electron cryo-microscopy (cryo-EM) reveals that the HIV stem-loop docks into the A site of the ribosome. Our results suggest that mRNA stem-loops can transiently escape the ribosome helicase by binding to the A site. Thus, the stem-loops can modulate gene expression by sterically hindering tRNA binding and inhibiting translation elongation. ribosome translation mRNA smFRET cryo-EM Medicine R Science Q Biology (General) Sarah Loerch verfasserin aut Clarence Ling verfasserin aut Andrei A Korostelev verfasserin aut Nikolaus Grigorieff verfasserin aut Dmitri N Ermolenko verfasserin aut In eLife eLife Sciences Publications Ltd, 2013 9(2020) (DE-627)728518384 (DE-600)2687154-3 2050084X nnns volume:9 year:2020 https://doi.org/10.7554/eLife.55799 kostenfrei https://doaj.org/article/716549eb2dca4eb8a1914d4e9a7819df kostenfrei https://elifesciences.org/articles/55799 kostenfrei https://doaj.org/toc/2050-084X Journal toc kostenfrei GBV_USEFLAG_A SYSFLAG_A GBV_DOAJ GBV_ILN_11 GBV_ILN_20 GBV_ILN_22 GBV_ILN_23 GBV_ILN_24 GBV_ILN_31 GBV_ILN_39 GBV_ILN_40 GBV_ILN_60 GBV_ILN_62 GBV_ILN_63 GBV_ILN_65 GBV_ILN_69 GBV_ILN_70 GBV_ILN_73 GBV_ILN_95 GBV_ILN_105 GBV_ILN_110 GBV_ILN_151 GBV_ILN_161 GBV_ILN_170 GBV_ILN_171 GBV_ILN_213 GBV_ILN_230 GBV_ILN_285 GBV_ILN_293 GBV_ILN_370 GBV_ILN_602 GBV_ILN_2003 GBV_ILN_2014 GBV_ILN_4012 GBV_ILN_4037 GBV_ILN_4112 GBV_ILN_4125 GBV_ILN_4126 GBV_ILN_4249 GBV_ILN_4305 GBV_ILN_4306 GBV_ILN_4307 GBV_ILN_4313 GBV_ILN_4322 GBV_ILN_4323 GBV_ILN_4324 GBV_ILN_4325 GBV_ILN_4335 GBV_ILN_4338 GBV_ILN_4367 GBV_ILN_4700 AR 9 2020
spelling	10.7554/eLife.55799 doi (DE-627)DOAJ004384385 (DE-599)DOAJ716549eb2dca4eb8a1914d4e9a7819df DE-627 ger DE-627 rakwb eng QH301-705.5 Chen Bao verfasserin aut mRNA stem-loops can pause the ribosome by hindering A-site tRNA binding 2020 Text txt rdacontent Computermedien c rdamedia Online-Ressource cr rdacarrier Although the elongating ribosome is an efficient helicase, certain mRNA stem-loop structures are known to impede ribosome movement along mRNA and stimulate programmed ribosome frameshifting via mechanisms that are not well understood. Using biochemical and single-molecule Förster resonance energy transfer (smFRET) experiments, we studied how frameshift-inducing stem-loops from E. coli dnaX mRNA and the gag-pol transcript of Human Immunodeficiency Virus (HIV) perturb translation elongation. We find that upon encountering the ribosome, the stem-loops strongly inhibit A-site tRNA binding and ribosome intersubunit rotation that accompanies translation elongation. Electron cryo-microscopy (cryo-EM) reveals that the HIV stem-loop docks into the A site of the ribosome. Our results suggest that mRNA stem-loops can transiently escape the ribosome helicase by binding to the A site. Thus, the stem-loops can modulate gene expression by sterically hindering tRNA binding and inhibiting translation elongation. ribosome translation mRNA smFRET cryo-EM Medicine R Science Q Biology (General) Sarah Loerch verfasserin aut Clarence Ling verfasserin aut Andrei A Korostelev verfasserin aut Nikolaus Grigorieff verfasserin aut Dmitri N Ermolenko verfasserin aut In eLife eLife Sciences Publications Ltd, 2013 9(2020) (DE-627)728518384 (DE-600)2687154-3 2050084X nnns volume:9 year:2020 https://doi.org/10.7554/eLife.55799 kostenfrei https://doaj.org/article/716549eb2dca4eb8a1914d4e9a7819df kostenfrei https://elifesciences.org/articles/55799 kostenfrei https://doaj.org/toc/2050-084X Journal toc kostenfrei GBV_USEFLAG_A SYSFLAG_A GBV_DOAJ GBV_ILN_11 GBV_ILN_20 GBV_ILN_22 GBV_ILN_23 GBV_ILN_24 GBV_ILN_31 GBV_ILN_39 GBV_ILN_40 GBV_ILN_60 GBV_ILN_62 GBV_ILN_63 GBV_ILN_65 GBV_ILN_69 GBV_ILN_70 GBV_ILN_73 GBV_ILN_95 GBV_ILN_105 GBV_ILN_110 GBV_ILN_151 GBV_ILN_161 GBV_ILN_170 GBV_ILN_171 GBV_ILN_213 GBV_ILN_230 GBV_ILN_285 GBV_ILN_293 GBV_ILN_370 GBV_ILN_602 GBV_ILN_2003 GBV_ILN_2014 GBV_ILN_4012 GBV_ILN_4037 GBV_ILN_4112 GBV_ILN_4125 GBV_ILN_4126 GBV_ILN_4249 GBV_ILN_4305 GBV_ILN_4306 GBV_ILN_4307 GBV_ILN_4313 GBV_ILN_4322 GBV_ILN_4323 GBV_ILN_4324 GBV_ILN_4325 GBV_ILN_4335 GBV_ILN_4338 GBV_ILN_4367 GBV_ILN_4700 AR 9 2020
allfields_unstemmed	10.7554/eLife.55799 doi (DE-627)DOAJ004384385 (DE-599)DOAJ716549eb2dca4eb8a1914d4e9a7819df DE-627 ger DE-627 rakwb eng QH301-705.5 Chen Bao verfasserin aut mRNA stem-loops can pause the ribosome by hindering A-site tRNA binding 2020 Text txt rdacontent Computermedien c rdamedia Online-Ressource cr rdacarrier Although the elongating ribosome is an efficient helicase, certain mRNA stem-loop structures are known to impede ribosome movement along mRNA and stimulate programmed ribosome frameshifting via mechanisms that are not well understood. Using biochemical and single-molecule Förster resonance energy transfer (smFRET) experiments, we studied how frameshift-inducing stem-loops from E. coli dnaX mRNA and the gag-pol transcript of Human Immunodeficiency Virus (HIV) perturb translation elongation. We find that upon encountering the ribosome, the stem-loops strongly inhibit A-site tRNA binding and ribosome intersubunit rotation that accompanies translation elongation. Electron cryo-microscopy (cryo-EM) reveals that the HIV stem-loop docks into the A site of the ribosome. Our results suggest that mRNA stem-loops can transiently escape the ribosome helicase by binding to the A site. Thus, the stem-loops can modulate gene expression by sterically hindering tRNA binding and inhibiting translation elongation. ribosome translation mRNA smFRET cryo-EM Medicine R Science Q Biology (General) Sarah Loerch verfasserin aut Clarence Ling verfasserin aut Andrei A Korostelev verfasserin aut Nikolaus Grigorieff verfasserin aut Dmitri N Ermolenko verfasserin aut In eLife eLife Sciences Publications Ltd, 2013 9(2020) (DE-627)728518384 (DE-600)2687154-3 2050084X nnns volume:9 year:2020 https://doi.org/10.7554/eLife.55799 kostenfrei https://doaj.org/article/716549eb2dca4eb8a1914d4e9a7819df kostenfrei https://elifesciences.org/articles/55799 kostenfrei https://doaj.org/toc/2050-084X Journal toc kostenfrei GBV_USEFLAG_A SYSFLAG_A GBV_DOAJ GBV_ILN_11 GBV_ILN_20 GBV_ILN_22 GBV_ILN_23 GBV_ILN_24 GBV_ILN_31 GBV_ILN_39 GBV_ILN_40 GBV_ILN_60 GBV_ILN_62 GBV_ILN_63 GBV_ILN_65 GBV_ILN_69 GBV_ILN_70 GBV_ILN_73 GBV_ILN_95 GBV_ILN_105 GBV_ILN_110 GBV_ILN_151 GBV_ILN_161 GBV_ILN_170 GBV_ILN_171 GBV_ILN_213 GBV_ILN_230 GBV_ILN_285 GBV_ILN_293 GBV_ILN_370 GBV_ILN_602 GBV_ILN_2003 GBV_ILN_2014 GBV_ILN_4012 GBV_ILN_4037 GBV_ILN_4112 GBV_ILN_4125 GBV_ILN_4126 GBV_ILN_4249 GBV_ILN_4305 GBV_ILN_4306 GBV_ILN_4307 GBV_ILN_4313 GBV_ILN_4322 GBV_ILN_4323 GBV_ILN_4324 GBV_ILN_4325 GBV_ILN_4335 GBV_ILN_4338 GBV_ILN_4367 GBV_ILN_4700 AR 9 2020
allfieldsGer	10.7554/eLife.55799 doi (DE-627)DOAJ004384385 (DE-599)DOAJ716549eb2dca4eb8a1914d4e9a7819df DE-627 ger DE-627 rakwb eng QH301-705.5 Chen Bao verfasserin aut mRNA stem-loops can pause the ribosome by hindering A-site tRNA binding 2020 Text txt rdacontent Computermedien c rdamedia Online-Ressource cr rdacarrier Although the elongating ribosome is an efficient helicase, certain mRNA stem-loop structures are known to impede ribosome movement along mRNA and stimulate programmed ribosome frameshifting via mechanisms that are not well understood. Using biochemical and single-molecule Förster resonance energy transfer (smFRET) experiments, we studied how frameshift-inducing stem-loops from E. coli dnaX mRNA and the gag-pol transcript of Human Immunodeficiency Virus (HIV) perturb translation elongation. We find that upon encountering the ribosome, the stem-loops strongly inhibit A-site tRNA binding and ribosome intersubunit rotation that accompanies translation elongation. Electron cryo-microscopy (cryo-EM) reveals that the HIV stem-loop docks into the A site of the ribosome. Our results suggest that mRNA stem-loops can transiently escape the ribosome helicase by binding to the A site. Thus, the stem-loops can modulate gene expression by sterically hindering tRNA binding and inhibiting translation elongation. ribosome translation mRNA smFRET cryo-EM Medicine R Science Q Biology (General) Sarah Loerch verfasserin aut Clarence Ling verfasserin aut Andrei A Korostelev verfasserin aut Nikolaus Grigorieff verfasserin aut Dmitri N Ermolenko verfasserin aut In eLife eLife Sciences Publications Ltd, 2013 9(2020) (DE-627)728518384 (DE-600)2687154-3 2050084X nnns volume:9 year:2020 https://doi.org/10.7554/eLife.55799 kostenfrei https://doaj.org/article/716549eb2dca4eb8a1914d4e9a7819df kostenfrei https://elifesciences.org/articles/55799 kostenfrei https://doaj.org/toc/2050-084X Journal toc kostenfrei GBV_USEFLAG_A SYSFLAG_A GBV_DOAJ GBV_ILN_11 GBV_ILN_20 GBV_ILN_22 GBV_ILN_23 GBV_ILN_24 GBV_ILN_31 GBV_ILN_39 GBV_ILN_40 GBV_ILN_60 GBV_ILN_62 GBV_ILN_63 GBV_ILN_65 GBV_ILN_69 GBV_ILN_70 GBV_ILN_73 GBV_ILN_95 GBV_ILN_105 GBV_ILN_110 GBV_ILN_151 GBV_ILN_161 GBV_ILN_170 GBV_ILN_171 GBV_ILN_213 GBV_ILN_230 GBV_ILN_285 GBV_ILN_293 GBV_ILN_370 GBV_ILN_602 GBV_ILN_2003 GBV_ILN_2014 GBV_ILN_4012 GBV_ILN_4037 GBV_ILN_4112 GBV_ILN_4125 GBV_ILN_4126 GBV_ILN_4249 GBV_ILN_4305 GBV_ILN_4306 GBV_ILN_4307 GBV_ILN_4313 GBV_ILN_4322 GBV_ILN_4323 GBV_ILN_4324 GBV_ILN_4325 GBV_ILN_4335 GBV_ILN_4338 GBV_ILN_4367 GBV_ILN_4700 AR 9 2020
allfieldsSound	10.7554/eLife.55799 doi (DE-627)DOAJ004384385 (DE-599)DOAJ716549eb2dca4eb8a1914d4e9a7819df DE-627 ger DE-627 rakwb eng QH301-705.5 Chen Bao verfasserin aut mRNA stem-loops can pause the ribosome by hindering A-site tRNA binding 2020 Text txt rdacontent Computermedien c rdamedia Online-Ressource cr rdacarrier Although the elongating ribosome is an efficient helicase, certain mRNA stem-loop structures are known to impede ribosome movement along mRNA and stimulate programmed ribosome frameshifting via mechanisms that are not well understood. Using biochemical and single-molecule Förster resonance energy transfer (smFRET) experiments, we studied how frameshift-inducing stem-loops from E. coli dnaX mRNA and the gag-pol transcript of Human Immunodeficiency Virus (HIV) perturb translation elongation. We find that upon encountering the ribosome, the stem-loops strongly inhibit A-site tRNA binding and ribosome intersubunit rotation that accompanies translation elongation. Electron cryo-microscopy (cryo-EM) reveals that the HIV stem-loop docks into the A site of the ribosome. Our results suggest that mRNA stem-loops can transiently escape the ribosome helicase by binding to the A site. Thus, the stem-loops can modulate gene expression by sterically hindering tRNA binding and inhibiting translation elongation. ribosome translation mRNA smFRET cryo-EM Medicine R Science Q Biology (General) Sarah Loerch verfasserin aut Clarence Ling verfasserin aut Andrei A Korostelev verfasserin aut Nikolaus Grigorieff verfasserin aut Dmitri N Ermolenko verfasserin aut In eLife eLife Sciences Publications Ltd, 2013 9(2020) (DE-627)728518384 (DE-600)2687154-3 2050084X nnns volume:9 year:2020 https://doi.org/10.7554/eLife.55799 kostenfrei https://doaj.org/article/716549eb2dca4eb8a1914d4e9a7819df kostenfrei https://elifesciences.org/articles/55799 kostenfrei https://doaj.org/toc/2050-084X Journal toc kostenfrei GBV_USEFLAG_A SYSFLAG_A GBV_DOAJ GBV_ILN_11 GBV_ILN_20 GBV_ILN_22 GBV_ILN_23 GBV_ILN_24 GBV_ILN_31 GBV_ILN_39 GBV_ILN_40 GBV_ILN_60 GBV_ILN_62 GBV_ILN_63 GBV_ILN_65 GBV_ILN_69 GBV_ILN_70 GBV_ILN_73 GBV_ILN_95 GBV_ILN_105 GBV_ILN_110 GBV_ILN_151 GBV_ILN_161 GBV_ILN_170 GBV_ILN_171 GBV_ILN_213 GBV_ILN_230 GBV_ILN_285 GBV_ILN_293 GBV_ILN_370 GBV_ILN_602 GBV_ILN_2003 GBV_ILN_2014 GBV_ILN_4012 GBV_ILN_4037 GBV_ILN_4112 GBV_ILN_4125 GBV_ILN_4126 GBV_ILN_4249 GBV_ILN_4305 GBV_ILN_4306 GBV_ILN_4307 GBV_ILN_4313 GBV_ILN_4322 GBV_ILN_4323 GBV_ILN_4324 GBV_ILN_4325 GBV_ILN_4335 GBV_ILN_4338 GBV_ILN_4367 GBV_ILN_4700 AR 9 2020
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container_title	eLife
authorswithroles_txt_mv	Chen Bao @@aut@@ Sarah Loerch @@aut@@ Clarence Ling @@aut@@ Andrei A Korostelev @@aut@@ Nikolaus Grigorieff @@aut@@ Dmitri N Ermolenko @@aut@@
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callnumber-first	Q - Science
author	Chen Bao
spellingShingle	Chen Bao misc QH301-705.5 misc ribosome misc translation misc mRNA misc smFRET misc cryo-EM misc Medicine misc R misc Science misc Q misc Biology (General) mRNA stem-loops can pause the ribosome by hindering A-site tRNA binding
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topic_title	QH301-705.5 mRNA stem-loops can pause the ribosome by hindering A-site tRNA binding ribosome translation mRNA smFRET cryo-EM
topic	misc QH301-705.5 misc ribosome misc translation misc mRNA misc smFRET misc cryo-EM misc Medicine misc R misc Science misc Q misc Biology (General)
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title	mRNA stem-loops can pause the ribosome by hindering A-site tRNA binding
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title_full	mRNA stem-loops can pause the ribosome by hindering A-site tRNA binding
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author_browse	Chen Bao Sarah Loerch Clarence Ling Andrei A Korostelev Nikolaus Grigorieff Dmitri N Ermolenko
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title_sort	mrna stem-loops can pause the ribosome by hindering a-site trna binding
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title_auth	mRNA stem-loops can pause the ribosome by hindering A-site tRNA binding
abstract	Although the elongating ribosome is an efficient helicase, certain mRNA stem-loop structures are known to impede ribosome movement along mRNA and stimulate programmed ribosome frameshifting via mechanisms that are not well understood. Using biochemical and single-molecule Förster resonance energy transfer (smFRET) experiments, we studied how frameshift-inducing stem-loops from E. coli dnaX mRNA and the gag-pol transcript of Human Immunodeficiency Virus (HIV) perturb translation elongation. We find that upon encountering the ribosome, the stem-loops strongly inhibit A-site tRNA binding and ribosome intersubunit rotation that accompanies translation elongation. Electron cryo-microscopy (cryo-EM) reveals that the HIV stem-loop docks into the A site of the ribosome. Our results suggest that mRNA stem-loops can transiently escape the ribosome helicase by binding to the A site. Thus, the stem-loops can modulate gene expression by sterically hindering tRNA binding and inhibiting translation elongation.
abstractGer	Although the elongating ribosome is an efficient helicase, certain mRNA stem-loop structures are known to impede ribosome movement along mRNA and stimulate programmed ribosome frameshifting via mechanisms that are not well understood. Using biochemical and single-molecule Förster resonance energy transfer (smFRET) experiments, we studied how frameshift-inducing stem-loops from E. coli dnaX mRNA and the gag-pol transcript of Human Immunodeficiency Virus (HIV) perturb translation elongation. We find that upon encountering the ribosome, the stem-loops strongly inhibit A-site tRNA binding and ribosome intersubunit rotation that accompanies translation elongation. Electron cryo-microscopy (cryo-EM) reveals that the HIV stem-loop docks into the A site of the ribosome. Our results suggest that mRNA stem-loops can transiently escape the ribosome helicase by binding to the A site. Thus, the stem-loops can modulate gene expression by sterically hindering tRNA binding and inhibiting translation elongation.
abstract_unstemmed	Although the elongating ribosome is an efficient helicase, certain mRNA stem-loop structures are known to impede ribosome movement along mRNA and stimulate programmed ribosome frameshifting via mechanisms that are not well understood. Using biochemical and single-molecule Förster resonance energy transfer (smFRET) experiments, we studied how frameshift-inducing stem-loops from E. coli dnaX mRNA and the gag-pol transcript of Human Immunodeficiency Virus (HIV) perturb translation elongation. We find that upon encountering the ribosome, the stem-loops strongly inhibit A-site tRNA binding and ribosome intersubunit rotation that accompanies translation elongation. Electron cryo-microscopy (cryo-EM) reveals that the HIV stem-loop docks into the A site of the ribosome. Our results suggest that mRNA stem-loops can transiently escape the ribosome helicase by binding to the A site. Thus, the stem-loops can modulate gene expression by sterically hindering tRNA binding and inhibiting translation elongation.
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title_short	mRNA stem-loops can pause the ribosome by hindering A-site tRNA binding
url	https://doi.org/10.7554/eLife.55799 https://doaj.org/article/716549eb2dca4eb8a1914d4e9a7819df https://elifesciences.org/articles/55799 https://doaj.org/toc/2050-084X
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