Activation of human beta-globin genes from nonerythroid cells by fusion with murine erythroleukemia cells.
AUTOR(ES)
Pyati, J
RESUMO
A human beta-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 beta-globin gene, human beta-globin mRNA was induced by Me2SO. Poly(A)-containing 10S human beta-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 beta-globin gene expression occurred only in hybrids that had retained human chromosome 11. Analysis of one hybrid bearing a deletion of both the beta-globin and lactate dehydrogenase A genes indicated that the beta-globin gene is located on the short arm of human chromosome 11. No other human chromosomes are required for human beta-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 with MEL cells provides a way to isolate permanent cell lines with functioning human globin genes. The technique should be useful for studying the biochemical basis for abnormal function of mutant globin genes, such as those present in individuals with the thalassemia syndromes.
ACESSO AO ARTIGO
http://www.pubmedcentral.nih.gov/articlerender.fcgi?artid=349631Documentos Relacionados
- High-level beta-globin expression after retroviral transfer of locus activation region-containing human beta-globin gene derivatives into murine erythroleukemia cells.
- Identification and characterization of a beta-globin promoter-binding factor from murine erythroleukemia cells.
- Expression of the human beta-globin gene after retroviral transfer into murine erythroleukemia cells and human BFU-E cells.
- A directly repeated sequence in the beta-globin promoter regulates transcription in murine erythroleukemia cells.
- DNA sequences involved in transcriptional regulation of the mouse beta-globin promoter in murine erythroleukemia cells.