Mono-ADP-ribosylation by ARTD10 restricts Chikungunya virus replication by interfering with the proteolytic activity of nsP2

Sarah Krieg; Fabian Pott; Laura Eckei; Maud Verheirstraeten; Mareike Bütepage; Barbara Lippok; Christine Goffinet; Bernhard Lüscher; Patricia Verheugd

MARylation of nsP2-459-798 is indicated by its mobility shift on SDS-PAGE (Fig. 5c ). The 307 inhibition of the protease function by ARTD10cat was dose dependent (Fig. 5d ). To interrogate 308 . CC-BY-NC-ND 4.0 International license author/funder. It is made available under a The copyright holder for this preprint (which was not peer-reviewed) is the . https://doi.org/10.1101/2020.01.07.896977 doi: bioRxiv preprint whether de-MARylation was sufficient to reactivate protease activity, we co-incubated the 309 substrate with nsP2-459-798, ARTD10cat and nsP3 or nsP3-macro as indicated (Fig. 5e) . MARylation, which was evident in the presence of hydrolase either by the reduced mobility 311 shift or by staining with the MAR reagent, reactivated protease activity (Fig. 5e ). The 312 processing efficiency was quantified by measuring the intensity of unprocessed substrate and 313 the two fragments by immunoblotting and densitometric analysis. This documented that 314