Fission yeast Cut2 required for anaphase has two destruction boxes. Academic Article uri icon

Overview

abstract

  • The fission yeast Schizosaccharomyces pombe cut2(+) gene is essential for sister chromatid separation. Cut2 protein, which locates in the interphase nucleus and along the metaphase spindle, disappears in anaphase with the same timing as mitotic cyclin destruction. This proteolysis depends on the APC (Anaphase-Promoting Complex)-cyclosome which contains ubiquitin ligase activity. The N-terminus of Cut2 contains two stretches similar to the mitotic cyclin destruction box. We show that both sequences (33RAPLGSTKQ and 52RTVLGGKST) serve as destruction boxes and are required for in vitro polyubiquitination and proteolysis. Cut2 with doubly mutated destruction boxes inhibits anaphase, whereas Cut2 with singly mutated boxes can suppress cut2 mutations. Strong expression of the N-terminal 73 residues containing the destruction boxes leads to the accumulation of endogenous cyclin and Cut2, and arrests cells in metaphase, whereas the same fragment with the mutated boxes does not. Cut2 proteolysis occurs in vitro using Xenopus mitotic extracts in the presence of functional destruction boxes. Furthermore, Cut2 is polyubiquitinated in an in vitro system using HeLa extracts, and this polyubiquitination requires the destruction boxes.

publication date

  • October 1, 1997

Research

keywords

  • Anaphase
  • Cell Cycle Proteins
  • Fungal Proteins
  • Schizosaccharomyces
  • Schizosaccharomyces pombe Proteins

Identity

PubMed Central ID

  • PMC1170228

Scopus Document Identifier

  • 0030824368

Digital Object Identifier (DOI)

  • 10.1093/emboj/16.19.5977

PubMed ID

  • 9312055

Additional Document Info

volume

  • 16

issue

  • 19