The genes for and regulation of the enzyme activities of two multifunctional proteins required for the de novo pathway for UMP biosynthesis in mammals.

Abstract

UMP biosynthesis requires six enzyme activities. Five of these enzyme centers are clustered into two multienzymatic proteins which are known to, or appear to, sequester the intermediates carbamyl approximately P, carbamyl aspartate and orotidylic acid. The advantages of sequestering these intermediates appear to be a conservation of energy, since two intermediates, carbamyl approximately P and orotidylate, might otherwise be rapidly degraded in mammalian cells. Carbamyl-aspartate appears not to be degraded rapidly in mammalian cells but it can pass into the blood and could possible disrupt brain metabolism by action as an acetylaspartate analog, if it passes the blood-brain barrier. For this, and possible for other reasons, there may be advantages to the fact that these intermediates are not other reasons, there may be advantages to the fact that these intermediates are not readily released from Complex A and U. In addition, these multienzymatic proteins may have other kinetic advantages, some of which have been discussed above. Studies with intact cells illustrate that azauridine, a chemical designed originally as an antineoplastic drug, produces a "ripple" effect when it inhibits the last enzyme of this pathway which leads to a sequential accumulation of pools of the various intermediates or their metabolites. This same agent increases the amount of some of the enzymes of this biosynthetic pathway in cells exposed to this drug. Both of these effects can negate the effectiveness of this potential antineoplastic drug. Sophisticated drug design may depend on whole-cell studies, such as those discussed here, in addition to the classic studies on the inhibition of a single enzyme center to select drugs that may be without significant side effects when they are finally tested in animals.