Multi-domain utilization by TUT4 and TUT7 in control of let-7 biogenesis

The uridyl transferases TUT4 and TUT7 switch between two modes of activity that either promotes let-7 expression (monoU) or marks it for degradation (oligoU). Lin28 modulates the switch via recruitment of TUT4(7) to pre-let-7 in stem cells and human cancers. We found TUT4(7) utilize two multi-domain functional modules during the switch from mono- to oligoU. The catalytic module (CM) is essential for both activities, while the Lin28-interacting module (LIM) is indispensible for oligoU. The TUT7 CM structure trapped in the monoU state, revealed a duplex RNA binding pocket that orients group II pre-let-7 hairpins to position the 1-nt overhang favor monoU addition. Conversely, the switch to oligoU requires the ZK domain of Lin28 to drive the formation of a stable ternary complex between pre-let-7 and the inactive LIM. Finally, ZK2 of TUT4(7) aids oligoU addition by engaging the growing oligoU tail through uracil-specific interactions.