Recombinant RNA-Dependent RNA Polymerase Complex of Ebola Virus.

ABSTRACT: Here we report on the expression, purification and characterization of recombinant ebola virus RNA-dependent RNA polymerase (EBOV RdRp). Active protein complexes composed of the large L protein and viral protein VP35 were isolated from insect cells and analyzed using a short primer/template substrate that allowed benchmarking against related enzymes. RNA synthesis by multiprotein complexes of EBOV, influenza B, respiratory syncytial virus (RSV) and monomeric enzymes of hepatitis C and Zika (ZIKV) viruses required a 5'-phosporylated primer. The minimum length of the primer varied between two and three nucleotides in this system. The EBOV enzyme utilizes Mg²⁺ as a co-factor and the D742A substitution provides an active site mutant that likely affects binding of the catalytic metal

SUBMITTER: Tchesnokov EP 

PROVIDER: S-EPMC5838098 | biostudies-literature | 2018 Mar

REPOSITORIES: biostudies-literature