Diagram of EBOV RNA editing and construction of EBOV GP mutants.
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Abstract
<p>(A) Schematic diagram of GP<sub>1,2</sub> and sGP. Membrane-bound GP<sub>1,2</sub> is encoded in the EBOV genome in two disjointed reading frames. The GP editing site is a tract of 7 A's approximately 900 nucleotides downstream of the start codon. Slippage of EBOV RNA-dependent RNA polymerase at the editing site results in insertion of an 8<sup>th</sup>-A which brings the two GP reading frames in register resulting in read-through translation of full-length membrane-bound trimeric GP<sub>1,2</sub>. Unedited transcripts contain a premature stop codon and produce truncated dimerized sGP. (B) EBOV GP and editing site mutants. Mutated nucleotides are shown in red and the primary gene products expressed by these constructs are also listed. (C) Expression of EBOV GP by wild type and mutant DNA constructs. HeLa cells were transfected with the wild type GP or editing site mutant constructs and GP expression was assayed by Western blot at 48 h post-transfection.</p
