The effect of 5-bromodeoxyuridine on DNA replication and cell division in Tetrahymena pyriformis

Populations of Tetrahymena pyriformis were grown in a chemically defined medium containing the thymidine analogue 5-bromodeoxyuridine (BUdR). About 65% of the thymidine sites in DNA were substituted by BUdR. During the first generation in the presence of BUdR, all DNA became hybrid. After the following cell division, in about 80% of the cells the second DNA replication round was initiated but no further cell division took place. The cells could be rescued by removing BUdR and adding thymidine. New replication took place before the first cell division. However, although the cells contained double heavy as well as hybrid DNA, only the hybrid DNA was replicated. After a full replication of the hybrid DNA, normal growth was restored. Melting profiles of normal, hybrid, and double heavy DNA indicated a structural change of the double heavy DNA.