TY - JOUR
T1 - Activation of human β-globin genes from nonerythroid cells by fusion with murine erythroleukemia cells
AU - Pyati, J.
AU - Kucherlapati, R. S.
AU - Skoultchi, A. I.
PY - 1980
Y1 - 1980
N2 - A human β-globin gene derived from an established human lymphoblast cell line was introduced into murine erythroleukemia (MEL) cells by cell fusion. The globin genes in MEL cells are inducible by dimethyl sulfoxide (Me2SO); induction leads to the accumulation of mouse globin mRNA and hemoglobin. Globin mRNA was not detected in the cytoplasm of the human lymphoblast cells, even at low levels, whether or not these cells were treated with Me2SO. In cell hybrids that had retained the lymphoblast-derived β-globin gene, human β-globin mRNA was induced by Me2SO. Poly(A)-containing 10S human β-globin mRNA was detected in the cytoplasm of the hybrid cells. Karyologic and isozymic analyses of a series of hybrids and subclones showed that human β-globin gene expression occurred only in hybrids that retained human chromosome 11. Analysis of one hybrid bearing a deletion of both the β-globin and lactate dehydrogenase A genes indicated that the β-globin gene is located on the short arm of human chromosome 11. No other human chromosomes are required for human β-globin gene expression in MEL cell hybrids. We conclude that the restricted expression of a globin gene in a human nonerythroid cell can be reversed. Furthermore, all components required for the transcription, processing, and transport to the cytoplasm of a human globin mRNA appear to be present in mouse erythroleukemia cells. Thus cell fusion wih MEL cells provides a way to isolate permanent cell lines with functioning human globin genes. The techniques should be useful for studying the biochemical basis for abnormal function of mutant globin genes, such as those present in individual with the thalassemia syndromes.
AB - A human β-globin gene derived from an established human lymphoblast cell line was introduced into murine erythroleukemia (MEL) cells by cell fusion. The globin genes in MEL cells are inducible by dimethyl sulfoxide (Me2SO); induction leads to the accumulation of mouse globin mRNA and hemoglobin. Globin mRNA was not detected in the cytoplasm of the human lymphoblast cells, even at low levels, whether or not these cells were treated with Me2SO. In cell hybrids that had retained the lymphoblast-derived β-globin gene, human β-globin mRNA was induced by Me2SO. Poly(A)-containing 10S human β-globin mRNA was detected in the cytoplasm of the hybrid cells. Karyologic and isozymic analyses of a series of hybrids and subclones showed that human β-globin gene expression occurred only in hybrids that retained human chromosome 11. Analysis of one hybrid bearing a deletion of both the β-globin and lactate dehydrogenase A genes indicated that the β-globin gene is located on the short arm of human chromosome 11. No other human chromosomes are required for human β-globin gene expression in MEL cell hybrids. We conclude that the restricted expression of a globin gene in a human nonerythroid cell can be reversed. Furthermore, all components required for the transcription, processing, and transport to the cytoplasm of a human globin mRNA appear to be present in mouse erythroleukemia cells. Thus cell fusion wih MEL cells provides a way to isolate permanent cell lines with functioning human globin genes. The techniques should be useful for studying the biochemical basis for abnormal function of mutant globin genes, such as those present in individual with the thalassemia syndromes.
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U2 - 10.1073/pnas.77.6.3435
DO - 10.1073/pnas.77.6.3435
M3 - Article
C2 - 6932030
AN - SCOPUS:0019307195
SN - 0027-8424
VL - 77
SP - 3435
EP - 3439
JO - Proceedings of the National Academy of Sciences of the United States of America
JF - Proceedings of the National Academy of Sciences of the United States of America
IS - 6 I
ER -