Proteomic analysis of the response to cell cycle arrests in human myeloid leukemia cells

Elife. 2015 Jan 2:4:e04534. doi: 10.7554/eLife.04534.

Abstract

Previously, we analyzed protein abundance changes across a 'minimally perturbed' cell cycle by using centrifugal elutriation to differentially enrich distinct cell cycle phases in human NB4 cells (Ly et al., 2014). In this study, we compare data from elutriated cells with NB4 cells arrested at comparable phases using serum starvation, hydroxyurea, or RO-3306. While elutriated and arrested cells have similar patterns of DNA content and cyclin expression, a large fraction of the proteome changes detected in arrested cells are found to reflect arrest-specific responses (i.e., starvation, DNA damage, CDK1 inhibition), rather than physiological cell cycle regulation. For example, we show most cells arrested in G2 by CDK1 inhibition express abnormally high levels of replication and origin licensing factors and are likely poised for genome re-replication. The protein data are available in the Encyclopedia of Proteome Dynamics (

Keywords: DNA damage; biochemistry; cell biology; cell cycle; human; mass spectrometry; proteomics; rereplication; serum starvation.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Cell Cycle Checkpoints* / drug effects
  • Cell Line, Tumor
  • Chromatin Assembly and Disassembly / drug effects
  • Culture Media, Serum-Free / pharmacology
  • Humans
  • Hydroxyurea / pharmacology
  • Leukemia, Myeloid, Acute / metabolism*
  • Leukemia, Myeloid, Acute / pathology
  • Proteomics / methods*
  • Quinolines / pharmacology
  • Signal Transduction / drug effects
  • Thiazoles / pharmacology

Substances

  • Culture Media, Serum-Free
  • Quinolines
  • RO 3306
  • Thiazoles
  • Hydroxyurea