Cargando…
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...
Autores principales: | , , , , , , , , , , , , , , , , , , , , |
---|---|
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 |
_version_ | 1783371520480903168 |
---|---|
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. |
format | Online Article Text |
id | pubmed-6239188 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2018 |
publisher | Cold Spring Harbor Laboratory Press |
record_format | MEDLINE/PubMed |
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 |
work_keys_str_mv | AT daoehan structureofthe30sribosomaldecodingcomplexatambienttemperature AT poitevinfrederic structureofthe30sribosomaldecodingcomplexatambienttemperature AT sierraraymondg structureofthe30sribosomaldecodingcomplexatambienttemperature AT gaticornelius structureofthe30sribosomaldecodingcomplexatambienttemperature AT raoyashas structureofthe30sribosomaldecodingcomplexatambienttemperature AT ciftcihalilibrahim structureofthe30sribosomaldecodingcomplexatambienttemperature AT aksitfulya structureofthe30sribosomaldecodingcomplexatambienttemperature AT mcgurkalex structureofthe30sribosomaldecodingcomplexatambienttemperature AT obrinskitrevor structureofthe30sribosomaldecodingcomplexatambienttemperature AT mgbampaul structureofthe30sribosomaldecodingcomplexatambienttemperature AT hayesbrandon structureofthe30sribosomaldecodingcomplexatambienttemperature AT delichtenbergcasper structureofthe30sribosomaldecodingcomplexatambienttemperature AT pardoavilafatima structureofthe30sribosomaldecodingcomplexatambienttemperature AT corsepiusnicholas structureofthe30sribosomaldecodingcomplexatambienttemperature AT zhanglindsey structureofthe30sribosomaldecodingcomplexatambienttemperature AT seabergmatthewh structureofthe30sribosomaldecodingcomplexatambienttemperature AT huntermarks structureofthe30sribosomaldecodingcomplexatambienttemperature AT liangmengling structureofthe30sribosomaldecodingcomplexatambienttemperature AT koglinjasone structureofthe30sribosomaldecodingcomplexatambienttemperature AT wakatsukisoichi structureofthe30sribosomaldecodingcomplexatambienttemperature AT demircihasan structureofthe30sribosomaldecodingcomplexatambienttemperature |