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

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

Open Access

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

Author(s) -

Joanna Sztuba-Solińska,

Larissa Diaz,

Mia R. Kumar,

Gaëlle Kolb,

Michael R. Wiley,

Jozwick Lucas,

Jens H. Kuhn,

Gustavo Palacios,

Sheli R. Radoshitzky,

Stuart F.J. Le Grice,

Reed F. Johnson

Publication year - 2016

Publication title -

nucleic acids research

Language(s) - English

Resource type - Journals

SCImago Journal Rank - 9.008

H-Index - 537

eISSN - 1362-4954

pISSN - 0305-1048

DOI - 10.1093/nar/gkw825

Subject(s) - biology , rna , virology , rna virus , microbiology and biotechnology , genetics , pseudoknot , viral replication , virus , gene

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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