Analysis of HeLa cell hypoxanthine phosphoribosyltransferase mutants and revertants by two-dimensional polyacrylamide gel electrophoresis: evidence for silent gene activation
| Autor: | Edmond Lee, Gregory Milman, Gurdev S. Ghangas, Jane R. McLaughlin, Matthew R. George |
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| Rok vydání: | 1976 |
| Předmět: |
Hypoxanthine Phosphoribosyltransferase
Multidisciplinary biology Isoelectric focusing cells Mutant genetic processes nutritional and metabolic diseases Molecular biology enzymes and coenzymes (carbohydrates) Epitopes Biochemistry Genes Hypoxanthine-guanine phosphoribosyltransferase Mutation biology.protein Phosphoribosyltransferase Missense mutation Electrophoresis Polyacrylamide Gel Polyacrylamide gel electrophoresis Gene HeLa Cells Research Article |
| Zdroj: | Proceedings of the National Academy of Sciences of the United States of America. 73(12) |
| ISSN: | 0027-8424 |
| Popis: | The spot corresponding to hypoxanthine phosphoribosyltransferase (HPRT; IMP:pyrophosphate phosphoribosyltransferase, EC 2.4.2.8) has been identified in two-dimensional polyacrylamide gels of HeLa cell extracts. This spot is absent in gels of 24 HPRT dificient mutants. A missense mutant displays a new HPRT spot at the same molecular weight but different isoelectric focusing position. Five independently isolated revertants of the missense mutant display spots corresponding to both the wild-type and mutant proteins indicating that they synthesize HPRT from two separate genes. If the missense protein is synthesized from a mutated form of the initially active HPRT gene, then wild-type HPRT protein in the revertants must be snythesized from a newly activated but prevously silent wild-type gene. The newly activated gene in the revertants of the missense mutation appears unstable producing a high frequency of spontaneous HPRT mutants. |
| Databáze: | OpenAIRE |
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