Few basepairing-independent motifs in the apical half of the avian HBV ε RNA stem-loop determine site-specific initiation of protein-priming

Abstract Hepadnaviruses, including human hepatitis B virus (HBV), replicate their tiny DNA genomes by protein-primed reverse transcription of a pregenomic (pg) RNA. Replication initiation as well as pgRNA encapsidation depend on the interaction of the viral polymerase, P protein, with the ε RNA elem...

Descripción completa

Guardado en:

Detalles Bibliográficos
Autores principales:	Markus Gajer, Katharina Dörnbrack, Christine Rösler, Bernadette Schmid, Jürgen Beck, Michael Nassal
Formato:	article
Lenguaje:	EN
Publicado:	Nature Portfolio 2017
Materias:	Medicine R Science Q
Acceso en línea:	https://doaj.org/article/7431dee15fa947bb97373f7fe4a05b75
Etiquetas:	Agregar Etiqueta Sin Etiquetas, Sea el primero en etiquetar este registro!

id	oai:doaj.org-article:7431dee15fa947bb97373f7fe4a05b75
record_format	dspace
spelling	oai:doaj.org-article:7431dee15fa947bb97373f7fe4a05b752021-12-02T15:05:48ZFew basepairing-independent motifs in the apical half of the avian HBV ε RNA stem-loop determine site-specific initiation of protein-priming10.1038/s41598-017-07657-z2045-2322https://doaj.org/article/7431dee15fa947bb97373f7fe4a05b752017-08-01T00:00:00Zhttps://doi.org/10.1038/s41598-017-07657-zhttps://doaj.org/toc/2045-2322Abstract Hepadnaviruses, including human hepatitis B virus (HBV), replicate their tiny DNA genomes by protein-primed reverse transcription of a pregenomic (pg) RNA. Replication initiation as well as pgRNA encapsidation depend on the interaction of the viral polymerase, P protein, with the ε RNA element, featuring a lower and an upper stem, a central bulge, and an apical loop. The bulge, somehow assisted by the loop, acts as template for a P protein-linked DNA oligo that primes full-length minus-strand DNA synthesis. Phylogenetic conservation and earlier mutational studies suggested the highly based-paired ε structure as crucial for productive interaction with P protein. Using the tractable duck HBV (DHBV) model we here interrogated the entire apical DHBV ε (Dε) half for sequence- and structure-dependent determinants of in vitro priming activity, replication, and, in part, in vivo infectivity. This revealed single-strandedness of the bulge, a following G residue plus the loop subsequence GUUGU as the few key determinants for priming and initiation site selection; unexpectedly, they functioned independently of a specific structure context. These data provide new mechanistic insights into avihepadnaviral replication initiation, and they imply a new concept towards a feasible in vitro priming system for human HBV.Markus GajerKatharina DörnbrackChristine RöslerBernadette SchmidJürgen BeckMichael NassalNature PortfolioarticleMedicineRScienceQENScientific Reports, Vol 7, Iss 1, Pp 1-16 (2017)
institution	DOAJ
collection	DOAJ
language	EN
topic	Medicine R Science Q
spellingShingle	Medicine R Science Q Markus Gajer Katharina Dörnbrack Christine Rösler Bernadette Schmid Jürgen Beck Michael Nassal Few basepairing-independent motifs in the apical half of the avian HBV ε RNA stem-loop determine site-specific initiation of protein-priming
description	Abstract Hepadnaviruses, including human hepatitis B virus (HBV), replicate their tiny DNA genomes by protein-primed reverse transcription of a pregenomic (pg) RNA. Replication initiation as well as pgRNA encapsidation depend on the interaction of the viral polymerase, P protein, with the ε RNA element, featuring a lower and an upper stem, a central bulge, and an apical loop. The bulge, somehow assisted by the loop, acts as template for a P protein-linked DNA oligo that primes full-length minus-strand DNA synthesis. Phylogenetic conservation and earlier mutational studies suggested the highly based-paired ε structure as crucial for productive interaction with P protein. Using the tractable duck HBV (DHBV) model we here interrogated the entire apical DHBV ε (Dε) half for sequence- and structure-dependent determinants of in vitro priming activity, replication, and, in part, in vivo infectivity. This revealed single-strandedness of the bulge, a following G residue plus the loop subsequence GUUGU as the few key determinants for priming and initiation site selection; unexpectedly, they functioned independently of a specific structure context. These data provide new mechanistic insights into avihepadnaviral replication initiation, and they imply a new concept towards a feasible in vitro priming system for human HBV.
format	article
author	Markus Gajer Katharina Dörnbrack Christine Rösler Bernadette Schmid Jürgen Beck Michael Nassal
author_facet	Markus Gajer Katharina Dörnbrack Christine Rösler Bernadette Schmid Jürgen Beck Michael Nassal
author_sort	Markus Gajer
title	Few basepairing-independent motifs in the apical half of the avian HBV ε RNA stem-loop determine site-specific initiation of protein-priming
title_short	Few basepairing-independent motifs in the apical half of the avian HBV ε RNA stem-loop determine site-specific initiation of protein-priming
title_full	Few basepairing-independent motifs in the apical half of the avian HBV ε RNA stem-loop determine site-specific initiation of protein-priming
title_fullStr	Few basepairing-independent motifs in the apical half of the avian HBV ε RNA stem-loop determine site-specific initiation of protein-priming
title_full_unstemmed	Few basepairing-independent motifs in the apical half of the avian HBV ε RNA stem-loop determine site-specific initiation of protein-priming
title_sort	few basepairing-independent motifs in the apical half of the avian hbv ε rna stem-loop determine site-specific initiation of protein-priming
publisher	Nature Portfolio
publishDate	2017
url	https://doaj.org/article/7431dee15fa947bb97373f7fe4a05b75
work_keys_str_mv	AT markusgajer fewbasepairingindependentmotifsintheapicalhalfoftheavianhbvernastemloopdeterminesitespecificinitiationofproteinpriming AT katharinadornbrack fewbasepairingindependentmotifsintheapicalhalfoftheavianhbvernastemloopdeterminesitespecificinitiationofproteinpriming AT christinerosler fewbasepairingindependentmotifsintheapicalhalfoftheavianhbvernastemloopdeterminesitespecificinitiationofproteinpriming AT bernadetteschmid fewbasepairingindependentmotifsintheapicalhalfoftheavianhbvernastemloopdeterminesitespecificinitiationofproteinpriming AT jurgenbeck fewbasepairingindependentmotifsintheapicalhalfoftheavianhbvernastemloopdeterminesitespecificinitiationofproteinpriming AT michaelnassal fewbasepairingindependentmotifsintheapicalhalfoftheavianhbvernastemloopdeterminesitespecificinitiationofproteinpriming
_version_	1718388711583907840

Few basepairing-independent motifs in the apical half of the avian HBV ε RNA stem-loop determine site-specific initiation of protein-priming

Ejemplares similares