Cargando…

Single-molecule FRET-Rosetta reveals RNA structural rearrangements during human telomerase catalysis

Maintenance of telomeres by telomerase permits continuous proliferation of rapidly dividing cells, including the majority of human cancers. Despite its direct biomedical significance, the architecture of the human telomerase complex remains unknown. Generating homogeneous telomerase samples has pres...

Descripción completa

Detalles Bibliográficos
Autores principales: Parks, Joseph W., Kappel, Kalli, Das, Rhiju, Stone, Michael D.
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Cold Spring Harbor Laboratory Press 2017
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC5238793/
https://www.ncbi.nlm.nih.gov/pubmed/28096444
http://dx.doi.org/10.1261/rna.058743.116
_version_ 1782495775651004416
author Parks, Joseph W.
Kappel, Kalli
Das, Rhiju
Stone, Michael D.
author_facet Parks, Joseph W.
Kappel, Kalli
Das, Rhiju
Stone, Michael D.
author_sort Parks, Joseph W.
collection PubMed
description Maintenance of telomeres by telomerase permits continuous proliferation of rapidly dividing cells, including the majority of human cancers. Despite its direct biomedical significance, the architecture of the human telomerase complex remains unknown. Generating homogeneous telomerase samples has presented a significant barrier to developing improved structural models. Here we pair single-molecule Förster resonance energy transfer (smFRET) measurements with Rosetta modeling to map the conformations of the essential telomerase RNA core domain within the active ribonucleoprotein. FRET-guided modeling places the essential pseudoknot fold distal to the active site on a protein surface comprising the C-terminal element, a domain that shares structural homology with canonical polymerase thumb domains. An independently solved medium-resolution structure of Tetrahymena telomerase provides a blind test of our modeling methodology and sheds light on the structural homology of this domain across diverse organisms. Our smFRET-Rosetta models reveal nanometer-scale rearrangements within the RNA core domain during catalysis. Taken together, our FRET data and pseudoatomic molecular models permit us to propose a possible mechanism for how RNA core domain rearrangement is coupled to template hybrid elongation.
format Online
Article
Text
id pubmed-5238793
institution National Center for Biotechnology Information
language English
publishDate 2017
publisher Cold Spring Harbor Laboratory Press
record_format MEDLINE/PubMed
spelling pubmed-52387932018-02-01 Single-molecule FRET-Rosetta reveals RNA structural rearrangements during human telomerase catalysis Parks, Joseph W. Kappel, Kalli Das, Rhiju Stone, Michael D. RNA Article Maintenance of telomeres by telomerase permits continuous proliferation of rapidly dividing cells, including the majority of human cancers. Despite its direct biomedical significance, the architecture of the human telomerase complex remains unknown. Generating homogeneous telomerase samples has presented a significant barrier to developing improved structural models. Here we pair single-molecule Förster resonance energy transfer (smFRET) measurements with Rosetta modeling to map the conformations of the essential telomerase RNA core domain within the active ribonucleoprotein. FRET-guided modeling places the essential pseudoknot fold distal to the active site on a protein surface comprising the C-terminal element, a domain that shares structural homology with canonical polymerase thumb domains. An independently solved medium-resolution structure of Tetrahymena telomerase provides a blind test of our modeling methodology and sheds light on the structural homology of this domain across diverse organisms. Our smFRET-Rosetta models reveal nanometer-scale rearrangements within the RNA core domain during catalysis. Taken together, our FRET data and pseudoatomic molecular models permit us to propose a possible mechanism for how RNA core domain rearrangement is coupled to template hybrid elongation. Cold Spring Harbor Laboratory Press 2017-02 /pmc/articles/PMC5238793/ /pubmed/28096444 http://dx.doi.org/10.1261/rna.058743.116 Text en © 2017 Parks et al.; Published by Cold Spring Harbor Laboratory Press for the RNA Society http://creativecommons.org/licenses/by-nc/4.0/ This article is distributed exclusively by the RNA Society for the first 12 months after the full-issue publication date (see http://rnajournal.cshlp.org/site/misc/terms.xhtml). After 12 months, it is available under a Creative Commons License (Attribution-NonCommercial 4.0 International), as described at http://creativecommons.org/licenses/by-nc/4.0/.
spellingShingle Article
Parks, Joseph W.
Kappel, Kalli
Das, Rhiju
Stone, Michael D.
Single-molecule FRET-Rosetta reveals RNA structural rearrangements during human telomerase catalysis
title Single-molecule FRET-Rosetta reveals RNA structural rearrangements during human telomerase catalysis
title_full Single-molecule FRET-Rosetta reveals RNA structural rearrangements during human telomerase catalysis
title_fullStr Single-molecule FRET-Rosetta reveals RNA structural rearrangements during human telomerase catalysis
title_full_unstemmed Single-molecule FRET-Rosetta reveals RNA structural rearrangements during human telomerase catalysis
title_short Single-molecule FRET-Rosetta reveals RNA structural rearrangements during human telomerase catalysis
title_sort single-molecule fret-rosetta reveals rna structural rearrangements during human telomerase catalysis
topic Article
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC5238793/
https://www.ncbi.nlm.nih.gov/pubmed/28096444
http://dx.doi.org/10.1261/rna.058743.116
work_keys_str_mv AT parksjosephw singlemoleculefretrosettarevealsrnastructuralrearrangementsduringhumantelomerasecatalysis
AT kappelkalli singlemoleculefretrosettarevealsrnastructuralrearrangementsduringhumantelomerasecatalysis
AT dasrhiju singlemoleculefretrosettarevealsrnastructuralrearrangementsduringhumantelomerasecatalysis
AT stonemichaeld singlemoleculefretrosettarevealsrnastructuralrearrangementsduringhumantelomerasecatalysis