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• W2003193311 abstract "The effects of media vitamin B12(CNB12), l-methionine, folic acid, dl-5-methyltetrahydrofolate (5-MeH4folate), homocysteine, and other nutrients on four one-carbon enzymes in cultured Chinese hamster ovary (CHO) cells were examined. Excess 10 mm methionine elevates the amount of B12 methyltransferase 1.8 – 2.3-fold at media folate concentrations of 0.2 – 2.0 μm. Conversely, excess 100 μm folic acid increases the amount of B12 holoenzyme by 2.4 – 3.0-fold when the medium contains 0.01 – 0.1 mm methionine. These increases in B12 methyltransferase promoted by 100 μm media folate and 10 mm methionine are inhibited by cycloheximide. 5-MeH4folate will support growth and induce methyltransferase synthesis more efficiently than folic acid. Upon transfer to methionine-free media, wild-type CHO cells will survive and can be repeatedly subcultured in the absence of exogenous methionine, provided it is supplemented with 1.0 μm CNB12, 0.1 mm homocysteine, and 100 μm folic acid or 10 μm dl-5-MeH4folate. No growth occurs if homocysteine is omitted, but a requirement for added CNB12 does not become evident until the cells have undergone at least two or three divisions. Survival upon transfer from 0.1 mm methionine-containing to methionine-free media is dependent upon the B12 holomethyltransferase content of the cells used as an inoculum. Inoculum cells must have been previously grown in media supplemented with 1.0 μm CNB12 to stabilize and convert apo- to holomethyltransferase, and 100 μm folate (or 10 μm dl-5-MeH4folate) to induce maximal enzyme-protein synthesis. Transfer to methionine-deficient medium does not result in more than a 20–25% increase in the cellular B12 enzyme content over the level already induced by 100 μm folate in 0.1 mm methionine-supplemented media. A mutant auxotroph CHO AUXB1 with a triple growth requirement for glycine + adenosine + thymidine (McBurney, M. W., and Whitmore, G. F. (1974) Cell, 2, 173) cannot survive in media lacking exogenous methionine. High concentrations of media folic acid or dl-5-MeH4folate fail to induce elevated amounts of B12 methyltransferase in this mutant. Excess 10 mm medium methionine does, however, elevate its B12 enzyme as in the parent CHO cells. An additional mutant AUXB3 that requires glycine + adenosine (McBurney, M. W., and Whitmore, G. F. (1974) Cell, 2, 173) barely survives in methionine-deficient media. It has a folate-induced B12 enzyme level intermediate between wild-type CHO cells and AUXB1. The level of B12 methyltransferase induced by high media folate concentrations is a critical determinant of CHO cell survival in methionine-free media." @default.
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• W2003193311 title "Folate-dependent enzymes in cultured Chinese hamster ovary cells: Induction of 5-methyltetrahydrofolate homocysteine cobalamin methyltransferase by folate and methionine" @default.
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• W2003193311 doi "https://doi.org/10.1016/0003-9861(75)90060-0" @default.
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