Immunological characterization of hypoxanthine-guanine phosphoribosyl transferase mutants of mouse L cells: Evidence for mutations at different loci in the HGPRT gene
- 1 April 1975
- journal article
- research article
- Published by Wiley in Journal of Cellular Physiology
- Vol. 85 (2), 307-320
- https://doi.org/10.1002/jcp.1040850217
Abstract
A large collection (105) of mouse L cell mutants lacking hypoxanthine‐guanine phosphoribosyl transferase activity (HGPRT; E. C. 2.4.2.8) were analyzed for the presence of serologically cross reacting material (CRM). Antibody directed against highly purified mouse liver HGPRT was used for detecting CRM activity by two methods: (1) the standard precipitation‐inhibition assay; and (2) a radioimmune‐precipitation assay. The latter assay proved to have far greater sensitivity for the detection of altered forms of HGPRT. Approximately 40% of the HGPRT− cell lines contain CRM activity (i.e., were CRM+). This indicates that a minimum of 40% of the HGPRT− clones arose as a result of mutations in the HGPRT structural gene. The CRM+ cell lines were shown to contain different levels of CRM activity. Measurements of the heat sensitivity of CRM in the different HGPRT− cell lines showed a broad spectrum of CRM heat inactivation kinetics. These latter two observations provide strong evidence that the mutations giving rise to the HGPRT−CRM+ phenotype occurred at different sites in the HGPRT structural gene.Keywords
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