title: Expression of human adenosine deaminase after fusion of adenosine deaminase-deficient cells with mouse fibroblasts.
abstract: Two human choriocarcinoma cell lines were shown to be deficient in adenosine deaminase (ADA; adenosine aminohydrolase, EC 3.5.4.4) such that they did not produce bands on starch gels after electrophoresis and histochemical staining. Radiometric assay indicated that their ADA specific activity was approximately 2% that of HeLa (human) cell controls. Subclone analysis of one of the lines indicated that this deficiency was representative of individual cells of the line. After fusion of these cells with mouse fibroblasts having high ADA activity, most independently isolated hybrid clones expressed one of two, or both, additional (to the mouse) bands of ADA activity after electrophoresis. The expression of these extra bands in hybrids was dependent upon actual fusion. The phenomenon was observed in 30 of 45 independently derived hybrid clones from four different fusion experiments involving two different parental lines from each species. The pattern of appearance of the extra bands in independent hybrid clones and the tendency of a hybrid clone to lose one of the extra bands through subsequent passages suggests that the bands were the products of human genetic material. The extra bands electrophoretically comigrated with human ADA 1 and 2 from human ADA-1-2 heterozygotes and the faster-migrating of the two extra bands comigrated with human ADA 1 from HeLa cells. Therefore, we suggest that the bands appearing in hybrids are the products of the 1 and 2 alleles of the human ADA locus. The human cells used for fusion were deficient in ADA activity but contained the genetic information for ADA 1 and 2. Fusion with mouse cells having ADA activity resulted in the activation of both human gene products coded for on separate homologous chromosomes. We conclude that the human ADA locus is under manipulatable genetic regulation.
mesh_terms: D000243:Adenosine Deaminase / Q000096:biosynthesis*; D000818:Animals; D002460:Cell Line; D005796:Genes; D006801:Humans; D006822:Hybrid Cells; D051379:Mice; D009700:Nucleoside Deaminases / Q000096:biosynthesis*
chemical_list: D009700:Nucleoside Deaminases; D000243:Adenosine Deaminase
references.citation: Am J Obstet Gynecol. 1965 Sep 15;93:266-8
references.pmid: 14336657
references.citation: Comp Biochem Physiol. 1965 Jan;14:101-20
references.pmid: 14288190
references.citation: Nature. 1964 Sep 12;203:1142-4
references.pmid: 14213660
references.citation: Science. 1964 Aug 14;145(3633):709-10
references.pmid: 14168277
references.citation: Exp Cell Res. 1964 Jan;33:19-28
references.pmid: 14109133
references.citation: J Biol Chem. 1951 Nov;193(1):265-75
references.pmid: 14907713
references.citation: Exp Cell Res. 1976 Dec;103(2):303-10
references.pmid: 137122
references.citation: Somatic Cell Genet. 1977 May;3(3):295-302
references.pmid: 305122
references.citation: Science. 1977 Feb 25;195(4280):783-5
references.pmid: 836588
references.citation: Lancet. 1972 Nov 18;2(7786):1067-9
references.pmid: 4117384
references.citation: Proc Natl Acad Sci U S A. 1976 Jan;73(1):213-7
references.pmid: 1061119
references.citation: Proc Natl Acad Sci U S A. 1974 Jan;71(1):45-9
references.pmid: 4129802
references.citation: Hum Hered. 1974;24(1):1-11
references.pmid: 4136545
references.citation: Chromosoma. 1974;47(2):147-56
references.pmid: 4140778
references.citation: Proc Natl Acad Sci U S A. 1967 Sep;58(3):1104-11
references.pmid: 5233838
references.citation: Nat New Biol. 1973 Dec 19;246(155):200-2
references.pmid: 4519601
references.citation: Proc Natl Acad Sci U S A. 1973 Sep;70(9):2590-4
references.pmid: 4354857
references.citation: J Cell Biol. 1974 Nov;63(2 Pt 1):505-14
references.pmid: 4371156
references.citation: Proc Natl Acad Sci U S A. 1971 Dec;68(12):3026-30
references.pmid: 4399833
references.citation: Proc Natl Acad Sci U S A. 1975 Jul;72(7):2682-6
references.pmid: 1058482
references.citation: Science. 1974 Sep 6;185(4154):859-62
references.pmid: 4367319
references.citation: Birth Defects Orig Artic Ser. 1975;11(1):124-7
references.pmid: 1148377
references.citation: J Cell Sci. 1975 May;17(3):567-78
references.pmid: 1170183
references.citation: Dev Biol. 1973 Apr;31(2):227-33
references.pmid: 4207016
references.citation: Proc Natl Acad Sci U S A. 1972 Nov;69(11):3273-7
references.pmid: 4508320
references.citation: Pediatr Res. 1976 Jul;10(7):642-6
references.pmid: 132630
references.citation: J Cell Sci. 1973 Nov;13(3):841-61
references.pmid: 4797867
references.citation: Science. 1965 Aug 27;149(3687):936-43
references.pmid: 5318249
references.citation: N Engl J Med. 1975 Apr 3;292(14):714-9
references.pmid: 1089883
references.citation: J Pediatr. 1975 Feb;86(2):169-81
references.pmid: 1089440
references.citation: Science. 1976 Jan 30;191(4225):392-4
references.pmid: 1246620
references.citation: Am J Hum Genet. 1975 Jan;27(1):46-52
references.pmid: 1155449
references.citation: Exp Cell Res. 1975 Jul;93(2):343-54
references.pmid: 1171770
references.citation: Ann Hum Genet. 1971 Oct;35(2):207-19
references.pmid: 5159535
references.citation: J Clin Endocrinol Metab. 1971 May;32(5):683-7
references.pmid: 5103722
references.citation: Biol Reprod. 1970 Apr;2(2):291-8
references.pmid: 5520327