Aberrant Replication Timing Induces Defective Chromosome Condensation in Drosophila ORC2 Mutants

Curr Biol. 2000 Dec 14-28;10(24):1547-56. doi: 10.1016/s0960-9822(00)00844-7.

Abstract

Background: The accurate duplication and packaging of the genome is an absolute prerequisite to the segregation of chromosomes in mitosis. To understand the process of cell-cycle chromosome dynamics further, we have performed the first detailed characterization of a mutation affecting mitotic chromosome condensation in a metazoan. Our combined genetic and cytological approaches in Drosophila complement and extend existing work employing yeast genetics and Xenopus in vitro extract systems to characterize higher-order chromosome structure and function.

Results: Two alleles of the ORC2 gene were found to cause death late in larval development, with defects in cell-cycle progression (delays in S-phase entry and metaphase exit) and chromosome condensation in mitosis. During S-phase progression in wild-type cells, euchromatin replicates early and heterochromatin replicates late. Both alleles disrupted the normal pattern of chromosomal replication, with some euchromatic regions replicating even later than heterochromatin. Mitotic chromosomes were irregularly condensed, with the abnormally late replicating regions of euchromatin exhibiting the greatest problems in mitotic condensation.

Conclusions: The results not only reveal novel functions for ORC2 in chromosome architecture in metazoans, they also suggest that the correct timing of DNA replication may be essential for the assembly of chromatin that is fully competent to undergo mitotic condensation.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Animals
  • Bromodeoxyuridine / pharmacology
  • Cell Cycle / physiology*
  • Cells, Cultured
  • Chromosomes / metabolism*
  • DNA / metabolism
  • DNA Replication* / drug effects
  • DNA-Binding Proteins / genetics
  • DNA-Binding Proteins / metabolism*
  • Drosophila Proteins
  • Drosophila melanogaster / embryology
  • Drosophila melanogaster / genetics
  • Drosophila melanogaster / physiology*
  • Female
  • In Situ Hybridization, Fluorescence
  • Microscopy, Fluorescence
  • Models, Biological
  • Mutation
  • Origin Recognition Complex
  • Phenotype
  • Time Factors

Substances

  • DNA-Binding Proteins
  • Drosophila Proteins
  • Orc2 protein, Drosophila
  • Origin Recognition Complex
  • DNA
  • Bromodeoxyuridine