The use of methylthioadenosine phosphorylase activity to select for human chromosome 9 in interspecies and intraspecies hybrid cells

Journal Article · Wed Sep 01 04:00:00 EDT 1993 · Somatic Cell and Molecular Genetics; (United States)

DOI:https://doi.org/10.1007/BF01233252· OSTI ID:5017418

Porterfield, B W; Pomykala, H; Maltepe, E; Bohlander, S K; Rowley, J D; Diaz, M O

Methylthioadenosine phosphorylase (MTAP) is an enzyme that functions in a salvage pathway for adenine synthesis. The locus that encodes MTAP activity has been mapped to human chromosome 9 (9q12-9pter) by analysis of mouse x human somatic cell hybrids. Cells that have MTAP activity will stop proliferating, and eventually die, in the presence of azaserine, an inhibitor of de novo purine synthesis, but can be rescued by the addition of methylthioadenosine (MTA) to the culture medium. Some mouse and human tumor cells lack MTAP activity and cannot be grown in the presence of azaserine and MTA. The authors fused MTAP competent human fibroblast cells to MTAP deficient mouse L-cells and selected for somatic cell hybrids, containing MTAP activity, in medium containing azaserine and MTA. In a separate experiment, a CHO cell x human fibroblast somatic cell hybrid, containing a normal copy of human chromosome 9, was used to prepare microcells, which were fused to an MTAP-deficient human leukemic cell line, CCRF-CEM. Somatic cell and microcell hybrids were shown to retain human chromosome 9 by fluorescence in situ hybridization using probes that hybridize to the interferon-alpha and -beta 1 genes on human chromosome 9 (9p21), and the centromere of human chromosome 9. This is the first report of complementation for MTAP activity being used to select for somatic cell hybrids and microcell hybrids that retain a human chromosome 9.

• DOE Contract Number: FG02-86ER60408
• OSTI ID: 5017418
• Journal Information: Somatic Cell and Molecular Genetics; (United States), Journal Name: Somatic Cell and Molecular Genetics; (United States) Vol. 19:5; ISSN 0740-7750; ISSN SCMGDN
• Country of Publication: United States
• Language: English

Related Subjects

550400* -- Genetics
59 BASIC BIOLOGICAL SCIENCES
ADENOSINE
ANIMAL CELLS
CHROMOSOMES
DETECTION
ENZYME ACTIVITY
ENZYMES
HUMAN CHROMOSOME 9
HUMAN CHROMOSOMES
HYBRIDIZATION
NUCLEOSIDES
NUCLEOTIDES
ORGANIC COMPOUNDS
PHOSPHORUS-GROUP TRANSFERASES
PHOSPHOTRANSFERASES
PROTEINS
RIBOSIDES
SOMATIC CELLS
TRANSFERASES