A small stem-loop structure of the Ebola virus trailer is essential for replication and interacts with heat-shock protein A8

Joanna Sztuba-Solinska, Larissa Diaz, Mia R. Kumar, Gaëlle Kolb, Michael R Wiley, Lucas Jozwick, Jens H. Kuhn, Gustavo Palacios, Sheli R. Radoshitzky, Stuart F.J. Le Grice, Reed F. Johnson

Research output: Contribution to journalArticle

7 Citations (Scopus)

Abstract

Ebola virus (EBOV) is a single-stranded negative-sense RNA virus belonging to the Filoviridae family. The leader and trailer non-coding regions of the EBOV genome likely regulate its transcription, replication, and progeny genome packaging. We investigated the cis-acting RNA signals involved in RNA-RNA and RNA-protein interactions that regulate replication of eGFP-encoding EBOV minigenomic RNA and identified heat shock cognate protein family A (HSC70) member 8 (HSPA8) as an EBOV trailer-interacting host protein. Mutational analysis of the trailer HSPA8 binding motif revealed that this interaction is essential for EBOV minigenome replication. Selective 2′-hydroxyl acylation analyzed by primer extension analysis of the secondary structure of the EBOV minigenomic RNA indicates formation of a small stem-loop composed of the HSPA8 motif, a 3′ stem-loop (nucleotides 1868-1890) that is similar to a previously identified structure in the replicative intermediate (RI) RNA and a panhandle domain involving a trailer-to-leader interaction. Results of minigenome assays and an EBOV reverse genetic system rescue support a role for both the panhandle domain and HSPA8 motif 1 in virus replication.

Original languageEnglish (US)
Pages (from-to)9831-9846
Number of pages16
JournalNucleic acids research
Volume44
Issue number20
DOIs
StatePublished - Jan 1 2016

Fingerprint

Ebolavirus
Heat-Shock Proteins
RNA
Virus Replication
Filoviridae
Genome
Reverse Genetics
Acylation
RNA Viruses
Product Packaging
Hydroxyl Radical
Proteins
Nucleotides

ASJC Scopus subject areas

  • Genetics

Cite this

A small stem-loop structure of the Ebola virus trailer is essential for replication and interacts with heat-shock protein A8. / Sztuba-Solinska, Joanna; Diaz, Larissa; Kumar, Mia R.; Kolb, Gaëlle; Wiley, Michael R; Jozwick, Lucas; Kuhn, Jens H.; Palacios, Gustavo; Radoshitzky, Sheli R.; Le Grice, Stuart F.J.; Johnson, Reed F.

In: Nucleic acids research, Vol. 44, No. 20, 01.01.2016, p. 9831-9846.

Research output: Contribution to journalArticle

Sztuba-Solinska, J, Diaz, L, Kumar, MR, Kolb, G, Wiley, MR, Jozwick, L, Kuhn, JH, Palacios, G, Radoshitzky, SR, Le Grice, SFJ & Johnson, RF 2016, 'A small stem-loop structure of the Ebola virus trailer is essential for replication and interacts with heat-shock protein A8', Nucleic acids research, vol. 44, no. 20, pp. 9831-9846. https://doi.org/10.1093/nar/gkw825
Sztuba-Solinska, Joanna ; Diaz, Larissa ; Kumar, Mia R. ; Kolb, Gaëlle ; Wiley, Michael R ; Jozwick, Lucas ; Kuhn, Jens H. ; Palacios, Gustavo ; Radoshitzky, Sheli R. ; Le Grice, Stuart F.J. ; Johnson, Reed F. / A small stem-loop structure of the Ebola virus trailer is essential for replication and interacts with heat-shock protein A8. In: Nucleic acids research. 2016 ; Vol. 44, No. 20. pp. 9831-9846.
@article{484c6afa510d4a3282d1bb35d6dd1d10,
title = "A small stem-loop structure of the Ebola virus trailer is essential for replication and interacts with heat-shock protein A8",
abstract = "Ebola virus (EBOV) is a single-stranded negative-sense RNA virus belonging to the Filoviridae family. The leader and trailer non-coding regions of the EBOV genome likely regulate its transcription, replication, and progeny genome packaging. We investigated the cis-acting RNA signals involved in RNA-RNA and RNA-protein interactions that regulate replication of eGFP-encoding EBOV minigenomic RNA and identified heat shock cognate protein family A (HSC70) member 8 (HSPA8) as an EBOV trailer-interacting host protein. Mutational analysis of the trailer HSPA8 binding motif revealed that this interaction is essential for EBOV minigenome replication. Selective 2′-hydroxyl acylation analyzed by primer extension analysis of the secondary structure of the EBOV minigenomic RNA indicates formation of a small stem-loop composed of the HSPA8 motif, a 3′ stem-loop (nucleotides 1868-1890) that is similar to a previously identified structure in the replicative intermediate (RI) RNA and a panhandle domain involving a trailer-to-leader interaction. Results of minigenome assays and an EBOV reverse genetic system rescue support a role for both the panhandle domain and HSPA8 motif 1 in virus replication.",
author = "Joanna Sztuba-Solinska and Larissa Diaz and Kumar, {Mia R.} and Ga{\"e}lle Kolb and Wiley, {Michael R} and Lucas Jozwick and Kuhn, {Jens H.} and Gustavo Palacios and Radoshitzky, {Sheli R.} and {Le Grice}, {Stuart F.J.} and Johnson, {Reed F.}",
year = "2016",
month = "1",
day = "1",
doi = "10.1093/nar/gkw825",
language = "English (US)",
volume = "44",
pages = "9831--9846",
journal = "Nucleic Acids Research",
issn = "0305-1048",
publisher = "Oxford University Press",
number = "20",

}

TY - JOUR

T1 - A small stem-loop structure of the Ebola virus trailer is essential for replication and interacts with heat-shock protein A8

AU - Sztuba-Solinska, Joanna

AU - Diaz, Larissa

AU - Kumar, Mia R.

AU - Kolb, Gaëlle

AU - Wiley, Michael R

AU - Jozwick, Lucas

AU - Kuhn, Jens H.

AU - Palacios, Gustavo

AU - Radoshitzky, Sheli R.

AU - Le Grice, Stuart F.J.

AU - Johnson, Reed F.

PY - 2016/1/1

Y1 - 2016/1/1

N2 - Ebola virus (EBOV) is a single-stranded negative-sense RNA virus belonging to the Filoviridae family. The leader and trailer non-coding regions of the EBOV genome likely regulate its transcription, replication, and progeny genome packaging. We investigated the cis-acting RNA signals involved in RNA-RNA and RNA-protein interactions that regulate replication of eGFP-encoding EBOV minigenomic RNA and identified heat shock cognate protein family A (HSC70) member 8 (HSPA8) as an EBOV trailer-interacting host protein. Mutational analysis of the trailer HSPA8 binding motif revealed that this interaction is essential for EBOV minigenome replication. Selective 2′-hydroxyl acylation analyzed by primer extension analysis of the secondary structure of the EBOV minigenomic RNA indicates formation of a small stem-loop composed of the HSPA8 motif, a 3′ stem-loop (nucleotides 1868-1890) that is similar to a previously identified structure in the replicative intermediate (RI) RNA and a panhandle domain involving a trailer-to-leader interaction. Results of minigenome assays and an EBOV reverse genetic system rescue support a role for both the panhandle domain and HSPA8 motif 1 in virus replication.

AB - Ebola virus (EBOV) is a single-stranded negative-sense RNA virus belonging to the Filoviridae family. The leader and trailer non-coding regions of the EBOV genome likely regulate its transcription, replication, and progeny genome packaging. We investigated the cis-acting RNA signals involved in RNA-RNA and RNA-protein interactions that regulate replication of eGFP-encoding EBOV minigenomic RNA and identified heat shock cognate protein family A (HSC70) member 8 (HSPA8) as an EBOV trailer-interacting host protein. Mutational analysis of the trailer HSPA8 binding motif revealed that this interaction is essential for EBOV minigenome replication. Selective 2′-hydroxyl acylation analyzed by primer extension analysis of the secondary structure of the EBOV minigenomic RNA indicates formation of a small stem-loop composed of the HSPA8 motif, a 3′ stem-loop (nucleotides 1868-1890) that is similar to a previously identified structure in the replicative intermediate (RI) RNA and a panhandle domain involving a trailer-to-leader interaction. Results of minigenome assays and an EBOV reverse genetic system rescue support a role for both the panhandle domain and HSPA8 motif 1 in virus replication.

UR - http://www.scopus.com/inward/record.url?scp=85016112037&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=85016112037&partnerID=8YFLogxK

U2 - 10.1093/nar/gkw825

DO - 10.1093/nar/gkw825

M3 - Article

VL - 44

SP - 9831

EP - 9846

JO - Nucleic Acids Research

JF - Nucleic Acids Research

SN - 0305-1048

IS - 20

ER -