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Structure of the 30S ribosomal decoding complex at ambient temperature

The ribosome translates nucleotide sequences of messenger RNA to proteins through selection of cognate transfer RNA according to the genetic code. To date, structural studies of ribosomal decoding complexes yielding high-resolution data have predominantly relied on experiments performed at cryogenic...

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Autores principales: Dao, E. Han, Poitevin, Frédéric, Sierra, Raymond G., Gati, Cornelius, Rao, Yashas, Ciftci, Halil Ibrahim, Akşit, Fulya, McGurk, Alex, Obrinski, Trevor, Mgbam, Paul, Hayes, Brandon, De Lichtenberg, Casper, Pardo-Avila, Fatima, Corsepius, Nicholas, Zhang, Lindsey, Seaberg, Matthew H., Hunter, Mark S., Liang, Mengling, Koglin, Jason E., Wakatsuki, Soichi, Demirci, Hasan
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Cold Spring Harbor Laboratory Press 2018
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC6239188/
https://www.ncbi.nlm.nih.gov/pubmed/30139800
http://dx.doi.org/10.1261/rna.067660.118
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author Dao, E. Han
Poitevin, Frédéric
Sierra, Raymond G.
Gati, Cornelius
Rao, Yashas
Ciftci, Halil Ibrahim
Akşit, Fulya
McGurk, Alex
Obrinski, Trevor
Mgbam, Paul
Hayes, Brandon
De Lichtenberg, Casper
Pardo-Avila, Fatima
Corsepius, Nicholas
Zhang, Lindsey
Seaberg, Matthew H.
Hunter, Mark S.
Liang, Mengling
Koglin, Jason E.
Wakatsuki, Soichi
Demirci, Hasan
author_facet Dao, E. Han
Poitevin, Frédéric
Sierra, Raymond G.
Gati, Cornelius
Rao, Yashas
Ciftci, Halil Ibrahim
Akşit, Fulya
McGurk, Alex
Obrinski, Trevor
Mgbam, Paul
Hayes, Brandon
De Lichtenberg, Casper
Pardo-Avila, Fatima
Corsepius, Nicholas
Zhang, Lindsey
Seaberg, Matthew H.
Hunter, Mark S.
Liang, Mengling
Koglin, Jason E.
Wakatsuki, Soichi
Demirci, Hasan
author_sort Dao, E. Han
collection PubMed
description The ribosome translates nucleotide sequences of messenger RNA to proteins through selection of cognate transfer RNA according to the genetic code. To date, structural studies of ribosomal decoding complexes yielding high-resolution data have predominantly relied on experiments performed at cryogenic temperatures. New light sources like the X-ray free electron laser (XFEL) have enabled data collection from macromolecular crystals at ambient temperature. Here, we report an X-ray crystal structure of the Thermus thermophilus 30S ribosomal subunit decoding complex to 3.45 Å resolution using data obtained at ambient temperature at the Linac Coherent Light Source (LCLS). We find that this ambient-temperature structure is largely consistent with existing cryogenic-temperature crystal structures, with key residues of the decoding complex exhibiting similar conformations, including adenosine residues 1492 and 1493. Minor variations were observed, namely an alternate conformation of cytosine 1397 near the mRNA channel and the A-site. Our serial crystallography experiment illustrates the amenability of ribosomal microcrystals to routine structural studies at ambient temperature, thus overcoming a long-standing experimental limitation to structural studies of RNA and RNA–protein complexes at near-physiological temperatures.
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spelling pubmed-62391882018-12-01 Structure of the 30S ribosomal decoding complex at ambient temperature Dao, E. Han Poitevin, Frédéric Sierra, Raymond G. Gati, Cornelius Rao, Yashas Ciftci, Halil Ibrahim Akşit, Fulya McGurk, Alex Obrinski, Trevor Mgbam, Paul Hayes, Brandon De Lichtenberg, Casper Pardo-Avila, Fatima Corsepius, Nicholas Zhang, Lindsey Seaberg, Matthew H. Hunter, Mark S. Liang, Mengling Koglin, Jason E. Wakatsuki, Soichi Demirci, Hasan RNA Report The ribosome translates nucleotide sequences of messenger RNA to proteins through selection of cognate transfer RNA according to the genetic code. To date, structural studies of ribosomal decoding complexes yielding high-resolution data have predominantly relied on experiments performed at cryogenic temperatures. New light sources like the X-ray free electron laser (XFEL) have enabled data collection from macromolecular crystals at ambient temperature. Here, we report an X-ray crystal structure of the Thermus thermophilus 30S ribosomal subunit decoding complex to 3.45 Å resolution using data obtained at ambient temperature at the Linac Coherent Light Source (LCLS). We find that this ambient-temperature structure is largely consistent with existing cryogenic-temperature crystal structures, with key residues of the decoding complex exhibiting similar conformations, including adenosine residues 1492 and 1493. Minor variations were observed, namely an alternate conformation of cytosine 1397 near the mRNA channel and the A-site. Our serial crystallography experiment illustrates the amenability of ribosomal microcrystals to routine structural studies at ambient temperature, thus overcoming a long-standing experimental limitation to structural studies of RNA and RNA–protein complexes at near-physiological temperatures. Cold Spring Harbor Laboratory Press 2018-12 /pmc/articles/PMC6239188/ /pubmed/30139800 http://dx.doi.org/10.1261/rna.067660.118 Text en © 2018 Dao et al.; Published by Cold Spring Harbor Laboratory Press for the RNA Society http://creativecommons.org/licenses/by/4.0/ This article, published in RNA, is available under a Creative Commons License (Attribution 4.0 International), as described at http://creativecommons.org/licenses/by/4.0/.
spellingShingle Report
Dao, E. Han
Poitevin, Frédéric
Sierra, Raymond G.
Gati, Cornelius
Rao, Yashas
Ciftci, Halil Ibrahim
Akşit, Fulya
McGurk, Alex
Obrinski, Trevor
Mgbam, Paul
Hayes, Brandon
De Lichtenberg, Casper
Pardo-Avila, Fatima
Corsepius, Nicholas
Zhang, Lindsey
Seaberg, Matthew H.
Hunter, Mark S.
Liang, Mengling
Koglin, Jason E.
Wakatsuki, Soichi
Demirci, Hasan
Structure of the 30S ribosomal decoding complex at ambient temperature
title Structure of the 30S ribosomal decoding complex at ambient temperature
title_full Structure of the 30S ribosomal decoding complex at ambient temperature
title_fullStr Structure of the 30S ribosomal decoding complex at ambient temperature
title_full_unstemmed Structure of the 30S ribosomal decoding complex at ambient temperature
title_short Structure of the 30S ribosomal decoding complex at ambient temperature
title_sort structure of the 30s ribosomal decoding complex at ambient temperature
topic Report
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC6239188/
https://www.ncbi.nlm.nih.gov/pubmed/30139800
http://dx.doi.org/10.1261/rna.067660.118
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