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

Gajer, Markus; Dörnbrack, Katharina; Rösler, Christine; Schmid, Bernadette; Beck, Jürgen; Nassal, Michael (2017). Few basepairing-independent motifs in the apical half of the avian HBV ε RNA stem-loop determine site-specific initiation of protein-priming. Scientific Reports, 7(1), p. 7120. Nature Publishing Group 10.1038/s41598-017-07657-z

[img]
Preview
Text
s41598-017-07657-z.pdf - Published Version
Available under License Creative Commons: Attribution (CC-BY).

Download (6MB) | Preview

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.

Item Type:

Journal Article (Original Article)

Division/Institute:

04 Faculty of Medicine > Department of Head Organs and Neurology (DKNS) > Clinic of Neurosurgery

UniBE Contributor:

Beck, Jürgen

Subjects:

600 Technology > 610 Medicine & health

ISSN:

2045-2322

Publisher:

Nature Publishing Group

Language:

English

Submitter:

Nicole Söll

Date Deposited:

01 Mar 2018 14:49

Last Modified:

26 Oct 2019 17:21

Publisher DOI:

10.1038/s41598-017-07657-z

PubMed ID:

28769080

BORIS DOI:

10.7892/boris.107936

URI:

https://boris.unibe.ch/id/eprint/107936

Actions (login required)

Edit item Edit item
Provide Feedback