AUUUA sequences compromise human insulin-like growth factor binding protein-1 mRNA stability

The instability of IGFBP-1 mRNA appears to play a role in regulating the expression of the IGFBP-1 gene, the 3' region of which contains five ATTTA sequences. We have studied the implication of these sequences for IGFBP-1 mRNA destabilization. Six plasmids were constructed, containing increasingly shorter lengths of IGFBP-1 cDNA, each with a successive ATTTA sequence deleted from the 3' end. These were stably transfected into two non-IGFBP-1-expressing (cervical carcinoma and neuroblastoma) cell lines. Kinetics studies following inhibition of transcription showed that (1) the half-life of the full-length messenger was 2.80 +/- 0.32 h; (2) deletion of each successive sequence (particularly the second and the fourth) yielded a transcript of increasing stability; and (3) the half-life of the AUUUA-free mRNA was 26.65 +/- 1.65 h. Although the primary source of IGFBP-1 is the liver, our results demonstrate that destabilization of its mRNA is not liver-specific. The ATTTA consensus sequences in the 3' untranslated region of the IGFBP-1 gene therefore provide a posttranscriptional regulation pathway that, combined with transcriptional regulation, may account for the variations in IGFBP-1 expression with developmental stage, nutritional status, and hormonal environment.