Structure of N-terminal domain of ZAP indicates how a zinc-finger protein recognizes complex RNA

Zinc-finger antiviral protein (ZAP) is a host factor that specifically inhibits the replication of certain viruses such as HIV-1 by targeting viral mRNA for degradation. Structural analysis now reveals a large RNA-binding surface comprising many positively charged residues and two cavities, thereby providing insight into how ZAP recognizes its target RNA. Zinc-finger antiviral protein (ZAP) is a host factor that specifically inhibits the replication of certain viruses, such as HIV-1, by targeting viral mRNA for degradation. How ZAP recognizes its target RNA has been unclear. Here we report the crystal structure of the N-terminal domain of rat ZAP (NZAP225), the major functional domain. The overall structure of NZAP225 resembles a tractor, with four zinc-finger motifs located at the bottom. Structural and functional analyses identified multiple positively charged residues and two putative RNA-binding cavities forming a large putative RNA-binding cleft. ZAP molecules interact to form a dimer that binds to a ZAP-responsive RNA molecule containing two ZAP-binding modules. These results provide insights into how ZAP binds specifically to complex target RNA.