Fig 1.

PABP binds specifically to the 3′ UTR of RCNMV RNA1. (A) Schematic representations of RCNMV genomic RNAs and STagT-fused 3′ UTRs of RNA1 and RNA2 (R1-3′ UTR-S and R2-3′ UTR-S). Bold lines indicate the virus-derived sequence of STagT-fused viral RNA fragments. (B) Silver-stained SDS-PAGE gel showing eluted STagT-RNA fragments and host proteins from a Strepto Tag affinity purification. The 72-kDa band in the left gel was excised, digested with trypsin, and subjected to LC-MS/MS. MM, molecular mass. The protein is NtPABP (AB673187). (C) STagT pulldown assay using R1-3′ UTR-S and R2-3′ UTR-S in BYLS20 expressing FLAG-tagged NtPABP. The rightmost lane represents a mock pulldown sample. The input lane contains 1.6% of the extract used for the STagT pulldown assay. Purified bait RNAs (top) were visualized using ethidium bromide (EtBr) fluorescence. NtPABP-FLAG (bottom) was detected by Western blotting using anti-FLAG M2 monoclonal antibody (Sigma-Aldrich).