Amino acid starvation affects the initiation frequency of nucleolar RNA polymerase.

Abstract

The synthesis of ribosomal precursor RNA in mouse ascites nucleoli derived from cells starved for amino acids is compared with the activity of nucleoli from control cells cultivated in the presence of all amino acids. It is shown that deprivation of a single essential amino acid from the culture medium results in a drastic decrease of the RNA-forming capacity of the isolated nucleoli by a factor of 2-3. This switchoff in rRNA synthesis is a very fast process. Half-maximal inactivation occurs after only 30 min. Addition of amino acids to starved cells leads to a rapid recovery, which is reflected by a sharp increase in the RNA polymerase activity of the isolated nucleoli. Studies on the molecular mechanism of this amino acid-mediated control of rRNA synthesis indicate that this effect is not caused by different growth rates of the RNA chains, but rather by an altered initiation frequency of the RNA polymerase in vivo. Whereas in nucleoli derived from cells grown in full medium almost all the polymerase is tightly bound in a transcriptional complex, a high amount of "free" polymerase which becomes active after addition of exogenous template is present in nucleoli from starved cells.