Orthogonal genome-wide screenings in bat cells identify MTHFD1 as a target of broad antiviral therapy

Danielle E Anderson; Jin Cui; Qian Ye; Baoying Huang; Wenhong Zu; Jing Gong; Weiqiang Liu; So Young Kim; Biao Guo Yan; Kristmundur Sigmundsson; Xiao Fang Lim; Fei Ye; Peihua Niu; Xuming Zhou; Wenjie Tan; Lin-Fa Wang; Xu Tan

MTHFD1 is a trifunctional enzyme involved in the one carbon (C1) metabolism pathway, which 28 is responsible for cellular production of purine, dTMP and methyl groups 29 . MTHFD1 has three 29 enzymatic functions including dehydrogenase and cyclohydrolase activities encoded in the N-30 terminal domain and formyl tetrahydrofolate (Formyl THF) synthetase activity encoded in the C-31 . CC-BY-NC 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 It is made available under a The copyright holder for this preprint (which was not peer-reviewed) is the .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. 03.29.014209 doi: bioRxiv preprint terminal domain ( Fig. 3a and 3b) . We suspected that MTHFD1 knockdown would lead to the 1 deficiency of cellular purine levels thereby reducing RNA replication. Indeed, supplementation of 2 purine analogs, either hypoxanthine or inosine in the media almost completely rescued the 3 replication of influenza A and Zika viruses in MTHFD1-knockout or knockdown PaKi cells as 4