Modulation of the Activity of an Avian Gene Transferred into a Mammalian Cell by Cell Fusion

Mouse A9 cells, deficient in hypoxanthine phosphoribosyltransferase (EC 2.4.2.8), were fused with normal chick erythrocytes and selected in hypoxanthine--aminopterin--thymidine medium for cells with hypoxanthine phosphoribosyltransferase activity. Recovered hybrid cells produced the chick hypoxanthi...

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Bibliographic Details
Published in:Proceedings of the National Academy of Sciences - PNAS 1974-04, Vol.71 (4), p.1398-1402
Main Authors: Klinger, Harold P., Shin, Seung-Il
Format: Article
Language:English
Subjects:
Aminopterin
Animal cells
Animals
Antiserum
Azaguanine
Biological Sciences: Genetics
Cell Fusion
Chick Embryo
Chromatography, Paper
Chromosomes
Clone Cells
Culture Media
Electrophoresis
Enzyme Induction
Enzyme Repression
Erythrocytes
Genes
Hybrid cells
Hybrid Cells - enzymology
Hybridity
Hypoxanthines
Hypoxia
L cells
Mice
Pentosyltransferases - analysis
Pentosyltransferases - biosynthesis
Precipitin Tests
Somatic cells
Species Specificity
Thymidine
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Summary:Mouse A9 cells, deficient in hypoxanthine phosphoribosyltransferase (EC 2.4.2.8), were fused with normal chick erythrocytes and selected in hypoxanthine--aminopterin--thymidine medium for cells with hypoxanthine phosphoribosyltransferase activity. Recovered hybrid cells produced the chick hypoxanthine phosphoribosyltransferase exclusively, as demonstrated by electrophoretic mobility and immunoprecipitation tests, even though no chick chromosomes or chick cell-surface antigens could be identified in the hybrids. Surprisingly, the expression of the chick hypoxanthine phosphoribosyltransferase activity in the mouse/chick hybrids required the presence of aminopterin in the growth medium; in its absence, enzyme synthesis decreased markedly. Because of the rapid and reversible modulation of hypoxanthine phosphoribosyltransferase activity, the hybrid cells could proliferate equally well in media containing hypoxanthine--aminopterin--thymidine or 8-azaguanine. Cellular selection was definitely ruled out as a possible cause. These results confirm previous reports that specific genetic information can be selectively transferred from one cell to another of a distant species. Furthermore, they demonstrate that an avian gene, whose activity is normally expressed constitutively, can become facultative when integrated into a mammalian cell. This seems to be the first instance where heterologous gene activity has been shown to be reversibly modulated in hybrid cells.
ISSN:0027-8424
1091-6490
DOI:10.1073/pnas.71.4.1398