The cyclin-dependent kinase inhibitor Dacapo promotes replication licensing during Drosophila endocycles

EMBO J. 2007 Apr 18;26(8):2071-82. doi: 10.1038/sj.emboj.7601648. Epub 2007 Mar 22.

Abstract

The endocycle is a developmentally programmed variant cell cycle in which cells undergo repeated rounds of DNA replication with no intervening mitosis. In Drosophila, the endocycle is driven by the oscillations of Cyclin E/Cdk2 activity. How the periodicity of Cyclin E/Cdk2 activity is achieved during endocycles is poorly understood. Here, we demonstrate that the p21(cip1)/p27(kip1)/p57(kip2)-like cyclin-dependent kinase inhibitor (CKI), Dacapo (Dap), promotes replication licensing during Drosophila endocycles by reinforcing low Cdk activity during the endocycle Gap-phase. In dap mutants, cells in the endocycle have reduced levels of the licensing factor Double Parked/Cdt1 (Dup/Cdt1), as well as decreased levels of chromatin-bound minichromosome maintenance (MCM2-7) complex. Moreover, mutations in dup/cdt1 dominantly enhance the dap phenotype in several polyploid cell types. Consistent with a reduced ability to complete genomic replication, dap mutants accumulate increased levels of DNA damage during the endocycle S-phase. Finally, genetic interaction studies suggest that dap functions to promote replication licensing in a subset of Drosophila mitotic cycles.

Publication types

  • Comparative Study
  • Research Support, N.I.H., Intramural
  • Research Support, Non-U.S. Gov't

MeSH terms

  • Animals
  • Bromodeoxyuridine
  • Cell Cycle / physiology*
  • Cyclin E / metabolism*
  • Cyclin-Dependent Kinase 2 / metabolism*
  • DNA Replication / physiology*
  • Drosophila / metabolism*
  • Drosophila Proteins / genetics
  • Drosophila Proteins / metabolism*
  • Flow Cytometry
  • Immunohistochemistry
  • Microscopy, Electron, Scanning
  • Mutation / genetics
  • Nuclear Proteins / genetics
  • Nuclear Proteins / metabolism*
  • Photoreceptor Cells, Invertebrate / ultrastructure

Substances

  • Cyclin E
  • Drosophila Proteins
  • Nuclear Proteins
  • dap protein, Drosophila
  • Cyclin-Dependent Kinase 2
  • cdk2 protein, Drosophila
  • Bromodeoxyuridine