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Octa-repeat domain of the mammalian prion protein mRNA forms stable A-helical hairpin structure rather than G-quadruplexes.
Sci Rep. 2019 02 21; 9(1):2465.SR

Abstract

Misfolding and aggregation of prion protein (PrP) causes neurodegenerative diseases like Creutzfeldt-Jakob disease (CJD) and scrapie. Besides the consensus that spontaneous conversion of normal cellular PrP[C] into misfolded and aggregating PrP[Sc] is the central event in prion disease, an alternative hypothesis suggests the generation of pathological PrP[Sc] by rare translational frameshifting events in the octa-repeat domain of the PrP mRNA. Ribosomal frameshifting most commonly relies on a slippery site and an adjacent stable RNA structure to stall translating ribosome. Hence, it is crucial to unravel the secondary structure of the octa-repeat domain of PrP mRNA. Each of the five octa-repeats contains a motif (GGCGGUGGUGGCUGGG) which alone in vitro forms a G-quadruplex. Since the propensity of mRNA to form secondary structure depends on the sequence context, we set to determine the structure of the complete octa-repeat region. We assessed the structure of full-length octa-repeat domain of PrP mRNA using dynamic light scattering (DLS), small angle X-ray scattering (SAXS), circular dichroism (CD) spectroscopy and selective 2'-hydroxyl acylation analysis by primer extension (SHAPE). Our data show that the PrP octa-repeat mRNA forms stable A-helical hairpins with no evidence of G-quadruplex structure even in the presence of G-quadruplex stabilizing agents.

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Authors+Show Affiliations

Czech A
Institute of Biochemistry and Molecular Biology University of Hamburg, Hamburg, Germany. andreas.czech@chemie.uni-hamburg.de.
Konarev PV
A. V. Shubnikov Institute of Crystallography of Federal Scientific Research Centre "Crystallography and Photonics" of Russian Academy of Sciences, Moscow, Russia. National Research Centre "Kurchatov Institute", Moscow, Russia.
Goebel I
Institute of Biochemistry and Molecular Biology University of Hamburg, Hamburg, Germany.
Svergun DI
European Molecular Biology Laboratory, Hamburg Outstation, c/o DESY, Hamburg, Germany.
Wills PR
Department of Physics, University of Auckland, Auckland, New Zealand.
Ignatova Z
Institute of Biochemistry and Molecular Biology University of Hamburg, Hamburg, Germany.

MeSH

Amino Acid SequenceCircular DichroismDynamic Light ScatteringG-QuadruplexesHeLa CellsHumansInverted Repeat SequencesMutationPrion ProteinsRNA, MessengerScattering, Small AngleX-Ray Diffraction

Pub Type(s)

Journal Article
Research Support, Non-U.S. Gov't

Language

eng

PubMed ID

30792490

Citation

Czech, Andreas, et al. "Octa-repeat Domain of the Mammalian Prion Protein mRNA Forms Stable A-helical Hairpin Structure Rather Than G-quadruplexes." Scientific Reports, vol. 9, no. 1, 2019, p. 2465.
Czech A, Konarev PV, Goebel I, et al. Octa-repeat domain of the mammalian prion protein mRNA forms stable A-helical hairpin structure rather than G-quadruplexes. Sci Rep. 2019;9(1):2465.
Czech, A., Konarev, P. V., Goebel, I., Svergun, D. I., Wills, P. R., & Ignatova, Z. (2019). Octa-repeat domain of the mammalian prion protein mRNA forms stable A-helical hairpin structure rather than G-quadruplexes. Scientific Reports, 9(1), 2465. https://doi.org/10.1038/s41598-019-39213-2
Czech A, et al. Octa-repeat Domain of the Mammalian Prion Protein mRNA Forms Stable A-helical Hairpin Structure Rather Than G-quadruplexes. Sci Rep. 2019 02 21;9(1):2465. PubMed PMID: 30792490.
* Article titles in AMA citation format should be in sentence-case
TY - JOUR T1 - Octa-repeat domain of the mammalian prion protein mRNA forms stable A-helical hairpin structure rather than G-quadruplexes. AU - Czech,Andreas, AU - Konarev,Petr V, AU - Goebel,Ingrid, AU - Svergun,Dmitri I, AU - Wills,Peter R, AU - Ignatova,Zoya, Y1 - 2019/02/21/ PY - 2018/05/21/received PY - 2018/12/19/accepted PY - 2019/2/23/entrez PY - 2019/2/23/pubmed PY - 2020/8/29/medline SP - 2465 EP - 2465 JF - Scientific reports JO - Sci Rep VL - 9 IS - 1 N2 - Misfolding and aggregation of prion protein (PrP) causes neurodegenerative diseases like Creutzfeldt-Jakob disease (CJD) and scrapie. Besides the consensus that spontaneous conversion of normal cellular PrP[C] into misfolded and aggregating PrP[Sc] is the central event in prion disease, an alternative hypothesis suggests the generation of pathological PrP[Sc] by rare translational frameshifting events in the octa-repeat domain of the PrP mRNA. Ribosomal frameshifting most commonly relies on a slippery site and an adjacent stable RNA structure to stall translating ribosome. Hence, it is crucial to unravel the secondary structure of the octa-repeat domain of PrP mRNA. Each of the five octa-repeats contains a motif (GGCGGUGGUGGCUGGG) which alone in vitro forms a G-quadruplex. Since the propensity of mRNA to form secondary structure depends on the sequence context, we set to determine the structure of the complete octa-repeat region. We assessed the structure of full-length octa-repeat domain of PrP mRNA using dynamic light scattering (DLS), small angle X-ray scattering (SAXS), circular dichroism (CD) spectroscopy and selective 2'-hydroxyl acylation analysis by primer extension (SHAPE). Our data show that the PrP octa-repeat mRNA forms stable A-helical hairpins with no evidence of G-quadruplex structure even in the presence of G-quadruplex stabilizing agents. SN - 2045-2322 UR - https://www.unboundmedicine.com/medline/citation/30792490/Octa_repeat_domain_of_the_mammalian_prion_protein_mRNA_forms_stable_A_helical_hairpin_structure_rather_than_G_quadruplexes_ DB - PRIME DP - Unbound Medicine ER -