Purification of the dihydrofolate reductases overproduced by 16 independently-derived antifolate-resistant sublines of Chinese hamster lung cells and their drug-sensitive parental cell line, DC-3F, has confirmed our original observation that two molecular weight classes of this enzyme, viz. Mr=21,000 and Mr=20,000, can be overproduced by drug-resistant cells. Of the 16 drug-resistant sublines analyzed, 12 overproduce the 21,000-dalton enzyme, while 4 overproduce the 20,000-dalton enzyme. Both molecular weight classes are found in the drug-sensitive parental line. Two-dimensional polyacrylamide gel electrophoresis of the two molecular weight classes of enzyme has shown that each is composed of two isoelectric forms, and that the relative distributions of the isoelectric forms within a molecular weight class are maintained during overproduction. In vitro translation experiments have demonstrated that the synthesis of these isoelectric forms and their relative distributions are mediated by mRNA. Finally, evidence is presented for the existence of two sets of three dihydrofolate reductase mRNAs in overproducing sublines. Both sets, within a subline, encode the same molecular weight class of dihydrofolate reductase, while each set directs the synthesis of only one of the isoelectric forms within that class. Although the genetic basis for these observations is not yet understood, it is clear that the production of dihydrofolate reductase by Chinese hamster lung cells is complex and probably involves the expression of multiple dihydrofolate reductase genes and/or alleles.
|Original language||English (US)|
|Number of pages||11|
|Journal||Journal of Biological Chemistry|
|State||Published - 1982|
All Science Journal Classification (ASJC) codes
- Molecular Biology
- Cell Biology