TPA-induced arrest of erythroid differentiation is coupled with downregulation of GATA-1 and upregulation of GATA-2 in an erythroid cell line SAM-1

Kamesaki, H; Michaud, Ginette Y.; Irving, Steven G.; Suwabe, Naruyoshi; Kamesaki, Saori; Okuma, Minoru; Cossman, Jeffrey

doi:10.1182/blood.v87.3.999.bloodjournal873999

Cited by 17 publications

(2 citation statements)

References 41 publications

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“…They contain GATA‐2 epitopes and thus appear to be GATA‐2 isoforms. In relation to this observation, GATA‐2 truncated forms have been found previously, but were attributed to degradation of the full‐length protein ( Wilson et al , 1990 ; Kamesaki et al , 1996 ). In our case, the integrity of extracts was established by monitoring other transcription factors, including GATA‐1.…”

Section: Discussionsupporting

confidence: 71%

Factor binding to the human γ‐globin gene distal CCAAT site: candidates for repression of the normal gene or activation of HPFH mutants

Partington

Patient

1998

Br J Haematol

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We have examined factor binding to the distal human gamma-globin CCAAT site and three naturally occurring hereditary persistence of fetal haemoglobin (HPFH) mutations of this site. Factor binding was examined using nuclear extracts from the erythroleukaemic cell lines K562 and MEL, and from A4 cells, a non-transformed mouse bone marrow stem cell line, using the electrophoretic mobility shift assay. Under standard binding conditions, in addition to the previously reported binding by a CCAAT factor (CP1) and GATA-1, the wild-type (wt) sequence bound high mobility factors which appeared to be GATA-2 isoforms. However, when the non-specific competitor conditions were varied, the binding profile with K562, but not MEL nuclear extract, was substantially altered. CP1 and GATA-1 were absent, and two new factors were detected, one of which bound preferentially to the Greek and Japanese non-deletion HPFH mutants. However, binding by the GATA-2 isoforms to the wt sequence was maintained with both cell types, as it was using the A4 cell line. With modified binding conditions, in A4 cells the two non-deletion and the Black deletion HPFH mutants each had a different protein binding profile which was lost on erythroid induction of the cells. We discuss the possibility that the GATA-2 isoforms bound to the wt sequence may function to suppress wt gamma gene expression in the bone marrow. Additionally, those factors which bind preferentially either to the deletion or non-deletion HPFH mutants may play positive roles in establishing an active chromatin structure.

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Section: Discussionsupporting

confidence: 71%

Factor binding to the human γ‐globin gene distal CCAAT site: candidates for repression of the normal gene or activation of HPFH mutants

Partington

Patient

1998

Br J Haematol

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“…Furthermore, in addition to GATA-1, GATA-2 is expressed in mammalian haematopoietic progenitor cells and the mRNA for GATA-2 declines as GATA-1 increases during erythroid differentiation (11). This finding has led to the suggestion that erythroid differentiation is regulated by a precise quantitative balance in the levels of GATA-1 and GATA-2 (12).…”

Section: Introductionmentioning

confidence: 99%

Phosphorylation of GATA-1 increases its DNA-binding affinity and is correlated with induction of human K562 erythroleukaemia cells

Partington

Patient

1999

Nucleic Acids Research

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We have investigated by electrophoretic mobility shift assay (EMSA) the level of GATA-1 DNA-binding activity in nuclear extracts prepared from the human erythroleukaemic cell line, K562, after erythroid induction by hemin, sodium butyrate (NaB) or Trichostatin A or treatment with N -acetylcysteine (NAC). Relative to extract from untreated cells, GATA-1 binding activity increased markedly in all cases. However, immunoblot analysis revealed unchanged levels of GATA-1 protein after induction. Incubation of induced but not uninduced K562 extracts with phosphatase prior to EMSA weakened the binding activity, suggesting that the increase in GATA-1 binding following induction of K562 cells was a consequence of phosphorylation. When the mouse erythroleukaemic cell line MEL was induced with dimethylsulphoxide (DMSO), NaB or NAC, GATA-1 binding activity fell with DMSO, rose significantly with NaB and remained at about the same level in NAC-induced cells. In this case immunoblotting revealed that GATA-1 protein levels were in accord with the EMSA data. The DNA-binding activities of induced and uninduced MEL cell nuclear extracts were decreased by incubation with phosphatase, showing that phosphoryl-ation and DNA binding of GATA-1 are already optimalin these cells. The DNA-binding activity of affinity-purified GATA-1 from MEL cells was also reduced by phosphatase treatment, showing that phosphorylation/dephosphorylation is directly affecting the factor. Furthermore, when a comparison was made by EMSA of nuclear extracts prepared from K562 and MEL cells untreated or incubated with okadaic acid, a phosphatase inhibitor, GATA-1 binding was seen to increase with K562 cells, whereas with MEL cells there was no change in GATA-1 binding. Overall the results suggest that the level of GATA-1 phosphorylation increases after the induction of K562, but not MEL cells, where GATA-1 is already highly phosphorylated. Furthermore, phosphorylation increases the binding affinity of GATA-1 for a canonical binding site.

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Transcription Factor GATA-2 Gene Is Located Near 3q21 Breakpoints in Myeloid Leukemia

Wieser

Volz

Vinatzer

et al. 2000

Biochemical and Biophysical Research Communications

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TPA-induced arrest of erythroid differentiation is coupled with downregulation of GATA-1 and upregulation of GATA-2 in an erythroid cell line SAM-1

Cited by 17 publications

References 41 publications

Factor binding to the human γ‐globin gene distal CCAAT site: candidates for repression of the normal gene or activation of HPFH mutants

Factor binding to the human γ‐globin gene distal CCAAT site: candidates for repression of the normal gene or activation of HPFH mutants

Phosphorylation of GATA-1 increases its DNA-binding affinity and is correlated with induction of human K562 erythroleukaemia cells

Transcription Factor GATA-2 Gene Is Located Near 3q21 Breakpoints in Myeloid Leukemia

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