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Isolation of Acid-soluble Nucleotides from Rat Tissues and Measurement of Their Specific Activity Following P³² Administration^*

R. Daoust and A. Cantero

( Montreal Cancer Institute, Notre-Dame Hospital, Montreal, Canada.)

1. A method was described to isolate free nucleotides from acid-soluble fractions of tissues.

2. This method was applied to normal rat liver, intestinal mucosa, and liver tumors, and several free nucleotides and derivatives were found to be present in appreciable amounts in these tissues.

3. The incorporation of radiophosphorus into free nucleotides was observed to be rapid and to proceed at similar rates in normal liver, intestinal mucosa and liver tumors.

These findings support the hypothesis that nucleic acids may be built up from a supply of preformed nucleotides. Furthermore, since the nucleic acids are known to incorporate radiophosphorus at different rates in these tissues, the radioactivity data suggest that the rate-limiting step in the biogenesis of nucleic acids is the formation of polynucleotides from individual nucleotides.

^* This work was supported by a grant from the Damon Runyon Memorial Fund for Cancer Research, Inc. A preliminary report was presented before the American Association for Cancer Research (8).

This investigation was conducted during tenure of a Damon Runyon Cancer Research Fellowship.

At present on leave at the Chester Beatty Research Institute, Royal Cancer Hospital, London, as Exchange Fellow of the British Empire Cancer Campaign.

Abbreviations are as follows: P³², radiophosphorus; A, adenine; G, guanine; U, uracil; T, thymine; C, cytosine; I, inosine; MP, monophosphate; DP, diphosphate; TP, triphosphate; DPN, diphosphopyridine nucleotide; TPN, triphosphopyridine nucleotide; RNA, ribonucleic acid; DNA deoxyribonucleic acid; TCA, trichloroacetic acid.

Received 7/14/55.