Unlinking of Cell Division from Deoxyribonucleic Acid Replication in a Temperature-sensitive Deoxyribonucleic Acid Synthesis Mutant of Escherichia coli

Abstract

A new type of temperature-sensitive deoxyribonucleic acid (DNA) synthesis mutant, which can divide without a completion of DNA replication, was isolated from a thymidine-requiring Escherichia coli strain by means of photo-bromouracil selection after nitrosoguanidine mutagenesis. In this mutant, in spite of the fact that DNA synthesis stopped immediately after the temperature shift from 30 to 41 C, cells could continue to divide, though at a reduced rate. This cell division without DNA synthesis at 41 C is further supported by the following results. (i) Cell division took place at high temperature without addition of thymidine but not at all at 30 C. The parent strain of the mutant did not divide at 41 C without thymidine. (ii) Smaller cells isolated from the culture grown at 41 C did not contain DNA. This was shown by chemical analysis of the smaller cells and on electron micrographs. Ability of cells to divide was examined according to sizes of cells. By using the culture at 30 C, cells of various sizes were separated by means of sucrose-density gradient centrifugation. It was found that all cell fractions, including the smallest one, could divide at high temperature. These results suggest that in this mutant the completion of DNA replication is not required for triggering cell division at high temperature. Heat sensitivity of a factor which links cell division with DNA replication appears to be responsible. Some possible mechanisms of the coordination between cell division and DNA replication are discussed.