Role of STAT3 and GATA-1 interactions in γ-globin gene expression

Xiao Yao, Sirisha Kodeboyina, Li Liu, James Dzandu, Jose Sangerman, Solomon F. Ofori-Acquah, Betty Sue Pace

Research output: Contribution to journalArticle

21 Citations (Scopus)

Abstract

Objective: We previously demonstrated a silencing role for signal transducers and activators of transcription 3 (STAT3) in γ-globin gene regulation in primary erythroid cells. Recently, GATA-1, a key transcription factor involved in hematopoietic cell development, was shown to directly inhibit STAT3 activity in vivo. Therefore, we completed studies to determine if interactions between these two factors influence γ-globin gene expression. Materials and Methods: Chromatin immunoprecipitation assay was used to ascertain in vivo protein binding at the γ-globin 5′ untranslated region (5′UTR); protein-protein interactions were examined by coimmunoprecipitation analysis. In vitro protein - DNA binding were completed using surface plasmon resonance and electrophoretic mobility shift assay. Activity of a luciferase γ-globin promoter reporter and levels of γ-globin messenger RNA and fetal hemoglobin in stable K562 cell lines overexpressing STAT3 and GATA-1, were used to determine the influence of the STAT3/GATA-1 interaction on γ-globin gene expression. Results: We observed interaction between STAT3 and GATA-1 in K562 and mouse erythroleukemia cells in vivo at the γ-globin 5′UTR by chromatin immunoprecipitation assay. Electrophoretic mobility shift assay performed with a 41-base pair γ-globin DNA probe (γ41) demonstrated the presence of STAT3 and GATA-1 proteins in complexes assembled at the γ-globin 5′UTR. A consensus STAT3 DNA probe inhibited GATA-1-binding in a concentration-dependent manner, and the converse was also true. Enforced STAT3 expression augmented its binding at the γ-globin 5′UTR in vivo and silenced γ-promoter-driven luciferase activity. Stable enforced STAT3 expression in K562 cells reduced endogenous γ-globin messenger RNA level. This effect was reversed by GATA-1. Conclusion: These data provide evidence that GATA-1 can reverse STAT3-mediated γ-globin gene silencing in erythroid cells.

Original languageEnglish (US)
Pages (from-to)889-900
Number of pages12
JournalExperimental Hematology
Volume37
Issue number8
DOIs
StatePublished - Aug 1 2009
Externally publishedYes

Fingerprint

STAT3 Transcription Factor
Globins
Gene Expression
5' Untranslated Regions
Erythroid Cells
K562 Cells
Chromatin Immunoprecipitation
DNA Probes
Electrophoretic Mobility Shift Assay
Luciferases
Fetal Hemoglobin
Leukemia, Erythroblastic, Acute
Messenger RNA
Proteins
Surface Plasmon Resonance
DNA-Binding Proteins
Gene Silencing
Protein Binding
Base Pairing
Transcription Factors

ASJC Scopus subject areas

  • Molecular Biology
  • Hematology
  • Genetics
  • Cell Biology
  • Cancer Research

Cite this

Yao, X., Kodeboyina, S., Liu, L., Dzandu, J., Sangerman, J., Ofori-Acquah, S. F., & Pace, B. S. (2009). Role of STAT3 and GATA-1 interactions in γ-globin gene expression. Experimental Hematology, 37(8), 889-900. https://doi.org/10.1016/j.exphem.2009.05.004

Role of STAT3 and GATA-1 interactions in γ-globin gene expression. / Yao, Xiao; Kodeboyina, Sirisha; Liu, Li; Dzandu, James; Sangerman, Jose; Ofori-Acquah, Solomon F.; Pace, Betty Sue.

In: Experimental Hematology, Vol. 37, No. 8, 01.08.2009, p. 889-900.

Research output: Contribution to journalArticle

Yao, X, Kodeboyina, S, Liu, L, Dzandu, J, Sangerman, J, Ofori-Acquah, SF & Pace, BS 2009, 'Role of STAT3 and GATA-1 interactions in γ-globin gene expression', Experimental Hematology, vol. 37, no. 8, pp. 889-900. https://doi.org/10.1016/j.exphem.2009.05.004
Yao X, Kodeboyina S, Liu L, Dzandu J, Sangerman J, Ofori-Acquah SF et al. Role of STAT3 and GATA-1 interactions in γ-globin gene expression. Experimental Hematology. 2009 Aug 1;37(8):889-900. https://doi.org/10.1016/j.exphem.2009.05.004
Yao, Xiao ; Kodeboyina, Sirisha ; Liu, Li ; Dzandu, James ; Sangerman, Jose ; Ofori-Acquah, Solomon F. ; Pace, Betty Sue. / Role of STAT3 and GATA-1 interactions in γ-globin gene expression. In: Experimental Hematology. 2009 ; Vol. 37, No. 8. pp. 889-900.
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