Initial function analysis of a novel erythroid differentiation related geneEDRF1 |
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Authors: | Duncheng Wang Yan Li Beifen Shen |
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Institution: | (1) Beijing Institute of Basic Medical Sciences, Academy of Military Medical Sciences, 100850 Beijing, China |
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Abstract: | Erythroid differentiation depends on the establishment of specific patterns of gene expression. Hypersensitive site 2 (HS2,
serving as a major enhancer of globin genes)-binding proteins may be involved in its natural open chromosomal environment
formation. Previously we prepared monoclonal antibodies against HS2-binding nuclear proteins of terminal differentiated erythroid
cells. By utilizing the monoclonal antibodies, we screened λ-gt11 human fetal liver cDNA expression library and obtained one
cDNA clone, which was named erythroid differentiation related gene (EDRF1, Genbank accession number AF040247), encompassing an entire open reading frame. We investigated the expression pattern ofEDRF1 by RT-PCR technique. And a clue to the function ofEDRF1 has been found from confirmation of high levels ofEDRF1 mRNA in differentiated K562 and human fetal liver tissue. To illuminate the function ofEDRF1 in K562 cells, sense and antisenseEDRF1 constructs were prepared and transfected into K562 cells. α-globin mRNA was down-regulated and EpoR (erythropoietin receptor)
mRNA expression was increased in antisense transfected cells. Cells transfected with sense construct grew more slowly than
control cells suggested by 3H] thimidine incorporation experiments. Suppression of K562 proliferation was accompanied by increased spontaneous hemoglobin
synthesis demonstrated by spectrometry. K562 cells transfected with sense construct exhibited reduced clongenicity compared
with control cells in methycellulose culture. These data provided the evidence thatEDRF1 can influence globin expression and hemoglobin synthesis in K562 cells and modulated self-renewal in K562 cells. |
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Keywords: | HS2 binding proteins hemopoiesis cell clongenicity expression pattern |
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