Differential Expression of Cloned Yeast Dihydrofolate Reductase Gene in <i>Escherichia coli</i>

While studying the expression of a cloned Saccharomyces cerevisiae dihydrofolate reductase gene in Escherichia coli, ' we observed that the yeast dihydrofolate reductase activity shows an inverse relationship with the growth rate of the organism? Thus, growth in rich medium causes a decrease and growth in minimal medium causes an increase in the yeast dihydrofolate reductase activity in yeast as well as in E. coli containing the cloned yeast DNA fragment.Since the observation of a sequence-specific binding of Lactobacillus casei dihydrofolate reductase to its own gene,) we are exploring the possibility that the yeast dihydrofolate reductase gene is under a negative control. Accordingly, a corepressor, most probably an end product, binds to dihydrofolate reductase and converts it to an active repressor molecule that forms the basis for autoregulation. (mdmg) 1. Broth 18 5.0 0.48 2. Yeast-nitrogen base 18 8.30 3. Broth 35 0.30 4. Yeast-nitrogen base 35 0.45 2.87 0.93 0.67 NATH et al.: DIHYDROFOLATE IN YEAST 127concentrations of 50 pg/ml is included in the growth medium at 3OoC, the dihydrofolate reductase activity remained the same as in TABLE 1. Methionine and adenine, two other end products of the dihydrofolate reductase reaction: are also unlikely candidates since they are the constituents present in the supplements. Of course, tetrahydrofolate, the immediate end product, is another likely candidate. We are in the process of purifying the enzyme from yeast and hope to use it in sequence-specific DNA binding studies9 to confirm the nature of the corepressor.

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Dihydrofolate Reductase in Yeast: Studies on Repression Mechanism^a

Nath¹,

Troise²,

Aldred³

1986

Annals of the New York Academy of Sciences

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Differential Expression of Cloned Yeast Dihydrofolate Reductase Gene in Escherichia coli

Cited by 1 publication

References 8 publications

Dihydrofolate Reductase in Yeast: Studies on Repression Mechanism^a

Dihydrofolate Reductase in Yeast: Studies on Repression Mechanism^a

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Differential Expression of Cloned Yeast Dihydrofolate Reductase Gene in Escherichia coli

Cited by 1 publication

References 8 publications

Dihydrofolate Reductase in Yeast: Studies on Repression Mechanisma

Dihydrofolate Reductase in Yeast: Studies on Repression Mechanisma

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Dihydrofolate Reductase in Yeast: Studies on Repression Mechanism^a

Dihydrofolate Reductase in Yeast: Studies on Repression Mechanism^a