Lin28-mediated control of let-7 microRNA expression by alternative TUTases Zcchc11 (TUT4) and Zcchc6 (TUT7)

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FIGURE 1.
FIGURE 1.

Domains of Zcchc11 required for Lin28-mediated pre-let-7 uridylation. (A) Schematic representation of Zcchc11 and truncations used for in vitro uridylation assays. (B) Uridylation assays with synthetic pre-let-7g carried out using Flag immunopurified (IP) Zcchc11 variants and IP Lin28. α-Flag Western blots show similar amounts of IP Zcchc11 within experiments. (C) Summary of Zcchc11 domain requirements from in vitro uridylation assays. (D) Reconstitution of Lin28-enhanced pre-let-7 uridylation with recombinant proteins. Zcchc11 truncation ΔPneumoG/C purified from either HEK293T (IP) or E. coli (Recombinant) was incubated with either Flag-Lin28 (IP) or 6x-His Lin28 (r.Lin28) in a uridylation assay with synthetic pre-let-7g. (Left) Schematic representation of the domains present in ΔPneumoG/C.

This Article

  1. Published in Advance August 16, 2012, doi: 10.1261/rna.034538.112 RNA 18: 1875-1885

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