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 18681890) 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 andHSPA8 motif 1 in virus replication.
Document Details
- Document Type
- Technical Report
- Publication Date
- Dec 02, 2016
- Accession Number
- AD1022217
Entities
People
- Gaelle Kolb
- Jens H. Kuhn
- Joanna Sztuba-solinska
- Larissa Diaz
- Lucas Jozwick
- Mia R. Kumar
- Michael R. Wiley
- Palacios Gustavo
- Reed F. Johnson
- Sheli Radoshitzky
- Stuart F. J. Le Grice
Organizations
- United States Army Medical Research Institute of Infectious Diseases