| Article ID | Journal | Published Year | Pages | File Type |
|---|---|---|---|---|
| 1946673 | Biochimica et Biophysica Acta (BBA) - Gene Regulatory Mechanisms | 2011 | 10 Pages |
Amino acid response (AAR) pathway is activated when cells are deprived of amino acids. In the present study, using the human colon cancer cell line SW480, we observed that DKK1, an antagonist of the Wnt pathway, was significantly induced at the mRNA level after the removal of amino acids from the medium. Addition of the amino alcohol histidinol, which prevents the formation of histidinyl-tRNAHis, also increased DKK1 mRNA to a level similar to that observed when cells were deprived of all amino acids. Transcriptional activity and stability of DKK1 mRNA were both increased in the amino acid-deprived condition. The induction of DKK1 gene expression was confirmed by the increased immunofluorescent staining of the DKK1 protein in the amino acid deprived condition. Although chromatin immunoprecipitation assays showed increased RNA Polymerase II binding at the DKK1 promoter in amino acid-limited conditions, ATF4 binding to the promoter is absent. Luciferase reporter assays did not detect any functional AARE within the DKK1 gene structure. Knockdown of ATF4 by siRNA did not affect the increase of DKK1 mRNA during amino acid limitation. Inhibition of ERK phosphorylation abolished the induction of DKK1. Our study revealed that DKK1 is a novel target gene in the response to amino acid deficiency and that the expression of DKK1 is up-regulated through an ATF4-independent and an ERK-dependent pathway.
Research highlights► The Wnt signaling pathway was affected by amino acid deprivation through the induction of DKK1. ► Transcriptional regulation of DKK1 by amino acid deficiency was by the phosphorylation of ERK. ► Knockdown of ATF4 does not inhibit the induction of DKK1 mRNA. ► The study demonstrated the interactions among Wnt, AAR, and MAPK/ERK signaling pathways in a colon cancer cell line. ► DKK1 is regulated by a novel amino acid response pathway that is independent of the master regulator ATF4.
