Cell-type-specific TEV protease cleavage reveals cohesin functions in Drosophila neurons

Dev Cell. 2008 Feb;14(2):239-51. doi: 10.1016/j.devcel.2007.12.009.

Abstract

Cohesin is a highly conserved multisubunit complex that holds sister chromatids together in mitotic cells. At the metaphase to anaphase transition, proteolytic cleavage of the alpha kleisin subunit (Rad21) by separase causes cohesin's dissociation from chromosomes and triggers sister-chromatid disjunction. To investigate cohesin's function in postmitotic cells, where it is widely expressed, we have created fruit flies whose Rad21 can be cleaved by TEV protease. Cleavage causes precocious separation of sister chromatids and massive chromosome missegregation in proliferating cells, but not disaggregation of polytene chromosomes in salivary glands. Crucially, cleavage in postmitotic neurons is lethal. In mushroom-body neurons, it causes defects in axon pruning, whereas in cholinergic neurons it causes highly abnormal larval locomotion. These data demonstrate essential roles for cohesin in nondividing cells and also introduce a powerful tool by which to investigate protein function in metazoa.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Animals
  • Axons / metabolism
  • Cell Cycle Proteins / metabolism*
  • Choline / metabolism
  • Chromatids / metabolism
  • Chromosomal Proteins, Non-Histone / metabolism*
  • Chromosome Segregation
  • Dendrites / metabolism
  • Drosophila Proteins / metabolism*
  • Drosophila melanogaster / cytology*
  • Drosophila melanogaster / embryology
  • Drosophila melanogaster / metabolism*
  • Embryo, Nonmammalian / cytology
  • Embryo, Nonmammalian / metabolism
  • Endopeptidases / metabolism*
  • Fertility
  • Larva
  • Locomotion
  • Mitosis
  • Mutation / genetics
  • Neurons / cytology*
  • Neurons / metabolism*
  • Nuclear Proteins / metabolism*
  • Organ Specificity
  • Protein Binding

Substances

  • Cell Cycle Proteins
  • Chromosomal Proteins, Non-Histone
  • Drosophila Proteins
  • Nuclear Proteins
  • cohesins
  • vtd protein, Drosophila
  • Endopeptidases
  • TEV protease
  • Choline