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

doi:10.1093/nar/gkw825

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 journal › Article

8 Scopus citations

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 language	English (US)
Pages (from-to)	9831-9846
Number of pages	16
Journal	Nucleic acids research
Volume	44
Issue number	20
DOIs	https://doi.org/10.1093/nar/gkw825
State	Published - 2016
Externally published	Yes

ASJC Scopus subject areas

Genetics

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10.1093/nar/gkw825

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Sztuba-Solinska, J., Diaz, L., Kumar, M. R., Kolb, G., Wiley, M. R., Jozwick, L., Kuhn, J. H., Palacios, G., Radoshitzky, S. R., Le Grice, S. F. J., & Johnson, R. F. (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, 44(20), 9831-9846. https://doi.org/10.1093/nar/gkw825

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, 2016, p. 9831-9846.

Research output: Contribution to journal › Article

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.

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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.",

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