Dok2 likely down-regulates Klf1 in mouse erythroleukemia cells

Anticancer Res. 2014 Aug;34(8):4561-7.

Abstract

Background/aim: Docking protein 2 (Dok2) is an adapter protein which is involved in hematopoiesis. However, it still remains unclear how Dok2 functions in regulation of transcription of hematopoietic genes. To address this issue, we knocked-down Dok2 mRNA in mouse erythroleukemia cells which highly express Dok2 intrinsically.

Materials and methods: Mouse erythroleukemia cells were transfected with Dok2 siRNA for 24 h and gene expression of erythroid differentiation-related genes, such as GATA binding protein 1 (Gata1), Krüppel-like factor 1 (Klf1), α-globin and β-globin were assessed by real-time polymerase chain reaction.

Results: Among the tested genes, expression of Klf1 exhibited a 1.94-fold increase when compared to the control 24 h after transfection. Immunocytochemistry and chromatin immunoprecipitation assays revealed that Dok2 protein localizes in the nucleus and binds to the promoter region of Klf1 gene.

Conclusion: Dok2 is able to control Klf1 expression by transcriptional regulation through directly binding to its promoter region.

Keywords: Dok2; KIf1; Mouse erythroleukemia cells; docking protein 2; erythropoiesis.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Adaptor Proteins, Signal Transducing / physiology*
  • Animals
  • Cell Line, Tumor
  • Down-Regulation
  • Gene Expression Regulation, Leukemic
  • Kruppel-Like Transcription Factors / genetics*
  • Leukemia, Erythroblastic, Acute / genetics*
  • Leukemia, Erythroblastic, Acute / pathology
  • Mice
  • Phosphoproteins / physiology*
  • Promoter Regions, Genetic

Substances

  • Adaptor Proteins, Signal Transducing
  • Dok2 protein, mouse
  • Kruppel-Like Transcription Factors
  • Phosphoproteins
  • erythroid Kruppel-like factor