Unknown

Dataset Information

0

An upstream open reading frame modulates ebola virus polymerase translation and virus replication.


ABSTRACT: Ebolaviruses, highly lethal zoonotic pathogens, possess longer genomes than most other non-segmented negative-strand RNA viruses due in part to long 5' and 3' untranslated regions (UTRs) present in the seven viral transcriptional units. To date, specific functions have not been assigned to these UTRs. With reporter assays, we demonstrated that the Zaire ebolavirus (EBOV) 5'-UTRs lack internal ribosomal entry site function. However, the 5'-UTRs do differentially regulate cap-dependent translation when placed upstream of a GFP reporter gene. Most dramatically, the 5'-UTR derived from the viral polymerase (L) mRNA strongly suppressed translation of GFP compared to a ?-actin 5'-UTR. The L 5'-UTR is one of four viral genes to possess upstream AUGs (uAUGs), and ablation of each uAUG enhanced translation of the primary ORF (pORF), most dramatically in the case of the L 5'-UTR. The L uAUG was sufficient to initiate translation, is surrounded by a "weak" Kozak sequence and suppressed pORF translation in a position-dependent manner. Under conditions where eIF2? was phosphorylated, the presence of the uORF maintained translation of the L pORF, indicating that the uORF modulates L translation in response to cellular stress. To directly address the role of the L uAUG in virus replication, a recombinant EBOV was generated in which the L uAUG was mutated to UCG. Strikingly, mutating two nucleotides outside of previously-defined protein coding and cis-acting regulatory sequences attenuated virus growth to titers 10-100-fold lower than a wild-type virus in Vero and A549 cells. The mutant virus also exhibited decreased viral RNA synthesis as early as 6 hours post-infection and enhanced sensitivity to the stress inducer thapsigargin. Cumulatively, these data identify novel mechanisms by which EBOV regulates its polymerase expression, demonstrate their relevance to virus replication and identify a potential therapeutic target.

SUBMITTER: Shabman RS 

PROVIDER: S-EPMC3561295 | biostudies-literature | 2013 Jan

REPOSITORIES: biostudies-literature

altmetric image

Publications

An upstream open reading frame modulates ebola virus polymerase translation and virus replication.

Shabman Reed S RS   Hoenen Thomas T   Groseth Allison A   Jabado Omar O   Binning Jennifer M JM   Amarasinghe Gaya K GK   Feldmann Heinz H   Basler Christopher F CF  

PLoS pathogens 20130131 1


Ebolaviruses, highly lethal zoonotic pathogens, possess longer genomes than most other non-segmented negative-strand RNA viruses due in part to long 5' and 3' untranslated regions (UTRs) present in the seven viral transcriptional units. To date, specific functions have not been assigned to these UTRs. With reporter assays, we demonstrated that the Zaire ebolavirus (EBOV) 5'-UTRs lack internal ribosomal entry site function. However, the 5'-UTRs do differentially regulate cap-dependent translation  ...[more]

Similar Datasets

2021-12-22 | GSE191262 | GEO
| S-EPMC137975 | biostudies-literature
2019-10-28 | GSE137021 | GEO
| S-EPMC6898956 | biostudies-literature
| S-EPMC7058706 | biostudies-literature
| S-EPMC8509022 | biostudies-literature
2022-09-28 | PXD030889 | Pride
| S-EPMC2652341 | biostudies-literature
| S-EPMC2603286 | biostudies-other
| S-EPMC9426467 | biostudies-literature