CUG binding protein 1 binds to a specific region within the human albumin 3′ untranslated region

3′ Untranslated regions (3′UTRs) of messenger RNAs have important roles in post-transcriptional regulation of gene expression and this is partly achieved through binding of specific proteins to sequences or structures within these regions. Previously, replacement of a native luciferase 3′UTR with the human albumin 3′UTR has been found to lead to a 10-fold increase in luciferase reporter activity. In this work we investigated protein binding to the human albumin 3′UTR. Electrophoretic mobility shift and UV cross-linking assays indicate that a ∼50 kDa protein from Chinese Hamster Ovary (CHO) cells binds to the albumin 3′UTR, and affinity experiments followed by proteomics identified this protein as CUG binding protein 1 (CUG-BP1, also known as CELF1). Deletion analysis of the albumin 3′UTR showed that nucleotides 1–50 and nucleotides 101–150 are not required for binding but that removal of nucleotides 51–100 caused a loss in binding. The results suggest that CUG-BP1 binds to nucleotides 51–100 of the human albumin 3′UTR. In human cells CUG-BP1 binding may thus play a role in regulation of albumin expression and, additionally, it may have a function in post-transcriptional control in CHO cells.

► A ∼50 kDa protein from CHO cells binds to the human albumin 3′UTR.
► Proteomics identified the protein as CUG binding protein 1 (CUG-BP1).
► siRNA knock-down of CUB-BP1 expression abolished binding.
► Nucleotides 51–100 of the 3′UTR play a particularly significant role in the binding.
► The results are consistent with a role for CUG-BP1 in post-transcriptional control.