Studies on endoreduplication. V. A three-dimensional scheme for diplo- and quadruple chromosomes and a model for DNA replication

Abstract

The metaphase appearance of quadruple chromosomes in colchicine-treated CHO cells was compared between air-dried and gently squashed preparations. A marked difference in morphology between the two methods suggested that the planar alignment of quadruple chromosomes is an artifact of the spreading process and that quadruple chromosomes are organized within the nucleus in a three-dimensional configuration. By analyzing the alignment of the original and replicated strands, using BrdU incorporation, the three-dimensional orientation of the chromatids in quadruple chromosomes could be traced. This analysis led to a new model for DNA replication. According to this model, an opening of a DNA base pair which rotates about 90 degrees with respect to the deoxyribose phosphate backbone precedes DNA replication, resulting in the formation of the newly replicated strands perpendicular to the original plane of the base pair. This model, although derived from endoreduplication, may be applicable to a general scheme for DNA replication during normal chromosome duplication.