Number OrnithineCitrulline Arginine

Note: + indicates growth; - indicates no growth.

medium supplemented only with ornithine. Finally, group III mutants grew only on medium supplemented with arginine.

Srb and Horowitz therefore proposed that the biochemical pathway leading to the amino acid arginine has at least three steps:

Step Step Step

1 2 3 precursor-* ornithine-* citrulline-* arginine tions in groups II and III affect steps that come after the production of ornithine. We've already established that group II mutations affect a step before the production of citrulline; so group II mutations must block the conversion of ornithine into citrulline.

Group Group

II III

mutations mutations ornithine-* citrulline-* arginine

Because group I mutations affect some step before the production of ornithine, we can conclude that they must affect the conversion of some precursor into ornithine. We can now outline the biochemical pathway yielding ornithine, citrulline, and arginine.

Group Group

I II

mutations mutations precursor-* ornithine-*

Group III

mutations citrulline-* arginine

They concluded that the mutations in group I affected step 1 of this pathway, mutations in group II affected step 2, and mutations in group III affected step 3. But how did they know that the order of the compounds in the biochemical pathway was correct?

Notice that, if step 1 is blocked by a mutation, then the addition of either ornithine or citrulline allows growth, because these compounds can still be converted into arginine (see Figure 15.3). Similarly, if step 2 is blocked, the addition of citrulline allows growth, but the addition of ornithine has no effect. If step 3 is blocked, the spores will grow only if arginine is added to the medium. The underlying principle is that an auxotrophic mutant cannot synthesize any compound that comes after the step blocked by a mutation.

Using this reasoning with the information in Table 15.1, we can see that the addition of arginine to the medium allows all three groups of mutants to grow. Therefore, biochemical steps affected by all the mutants precede the step that results in arginine. The addition of citrulline allows group I and group II mutants to grow but not group III mutants; therefore, group III mutations must affect a biochemical step that takes place after the production of citrulline but before the production of arginine.

Group

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