Evidence that Poly(A) Binding Protein C1 Binds Nuclear Pre-mRNA Poly(A) Tails

Abstract

In mammalian cells, poly(A) binding protein C1 (PABP C1) has well-known roles in mRNA translation and decay in the cytoplasm. However, PABPC1 also shuttles in and out of the nucleus, and its nuclear function is unknown. Here, we show that PABPC1, like the major nuclear poly(A) binding protein PABPN1, associates with nuclear pre-mRNAs that are polyadenylated and intron containing. PABPC1 does not bind nonpolyadenylated histone mRNA, indicating that the interaction of PABPC1 with pre-mRNA requires a poly(A) tail. Consistent with this conclusion, UV cross-linking results obtained using intact cells reveal that PABPC1 binds directly to pre-mRNA poly(A) tails in vivo. We also show that PABPC1 immunopurifies with poly(A) polymerase, suggesting that PABPC1 is acquired by polyadenylated transcripts during poly(A) tail synthesis. Our findings demonstrate that PABPC1 associates with polyadenylated transcripts earlier in mammalian mRNA biogenesis than previously thought and offer insights into the mechanism by which PABPC1 is recruited to newly synthesized poly(A). Our results are discussed in the context of pre-mRNA processing and stability and mRNA trafficking and the pioneer round of translation.

We thank Rick Lloyd for anti-PABPC1, Luiz Penalva and Jack Keene for pSHREKK-PABPC1, Dave Bear for anti-PABPN1, Guy Rouleau for pEGFP-C2-PABPN1, Anders Virtanen for anti-PAP-γ, Yi-Tao Yu and Allan Jacobson for useful information, and Dave Bear and Holly Kuzmiak for helpful conversations and comments on the manuscript.

This work was supported by NIH R01 GM59614 to L.E.M. N.H. was supported in part by a fellowship from the Japan Society for the Promotion of Science.