Localization of macromolecules in Escherichia coli. I. DNA and proteins

Abstract

If thin sections of Escherichia coli, labeled uniformly with tritium, are radioautographed calculations, based on the distribution of section sizes show that the number of H(3) decays per section should be very close to a Poisson distribution. We might, therefore, expect that the distribution of radioautographic grain counts among random cross-sections should follow a Poisson distribution. It can then be inferred that a deviation from a Poisson indicates a high concentration of label in a preferred region. This region can then be identified by analysis of serial section and comparison with electron micrographs. Sections of cells labeled with leucine-H3 gave a Poisson distribution of grain counts, and it was concluded that proteins were distributed fairly uniformly throughout the cell. The situation was not changed if labeled cells were placed in chloramphenicol or if very short pulses of label were used. When Escherichia coli is grown in presence of chloramphenicol a major morphological change concerns the nuclear region: it becomes more regular in outline, nearly spherical, and occupies a smaller proportion of the cell length. The previously described association between DNA labeled with thymidine-H3 and the nuclear region was confirmed by showing that the distribution of the label in the cell followed exactly the morphological changes of the nuclear region. It was also shown that the concentration of DNA in the nuclear region was at least 45 times higher than that of the cytoplasm. Several morphological features of cells grown in chloramphenicol and examined in the electron microscope are discussed.