Microtubule interfering agents and KSP inhibitors induce the phosphorylation of the nuclear protein p54(nrb), an event linked to G2/M arrest

J Proteomics. 2009 Jan 30;71(6):592-600. doi: 10.1016/j.jprot.2008.09.001. Epub 2008 Sep 14.

Abstract

Microtubule interfering agents (MIAs) are anti-tumor drugs that inhibit microtubule dynamics, while kinesin spindle protein (KSP) inhibitors are substances that block the formation of the bipolar spindle during mitosis. All these compounds cause G2/M arrest and cell death. Using 2D-PAGE followed by Nano-LC-ESI-Q-ToF analysis, we found that MIAs such as vincristine (Oncovin) or paclitaxel (Taxol) and KSP inhibitors such as S-tritil-l-cysteine induce the phosphorylation of the nuclear protein p54(nrb) in HeLa cells. Furthermore, we demonstrate that cisplatin (Platinol), an anti-tumor drug that does not cause M arrest, does not induce this modification. We show that the G2/M arrest induced by MIAs is required for p54(nrb) phosphorylation. Finally, we demonstrate that CDK activity is required for MIA-induced phosphorylation of p54(nrb).

Publication types

  • Research Support, N.I.H., Extramural
  • Research Support, Non-U.S. Gov't

MeSH terms

  • Antineoplastic Agents, Phytogenic / pharmacology*
  • Cell Line
  • Cisplatin / pharmacology
  • Cysteine / analogs & derivatives
  • Cysteine / pharmacology
  • DNA-Binding Proteins
  • Electrophoresis, Gel, Two-Dimensional
  • G2 Phase / drug effects
  • G2 Phase / physiology*
  • Humans
  • Kinesins / antagonists & inhibitors
  • Kinesins / metabolism*
  • Microtubules
  • Mitosis / physiology*
  • Mitosis Modulators / pharmacology
  • Nuclear Matrix-Associated Proteins / metabolism*
  • Octamer Transcription Factors / metabolism*
  • Paclitaxel / pharmacology
  • Phosphorylation
  • RNA-Binding Proteins / metabolism*
  • Tandem Mass Spectrometry
  • Tubulin Modulators / pharmacology*
  • Vincristine / pharmacology

Substances

  • Antineoplastic Agents, Phytogenic
  • DNA-Binding Proteins
  • KIF11 protein, human
  • Mitosis Modulators
  • NONO protein, human
  • Nuclear Matrix-Associated Proteins
  • Octamer Transcription Factors
  • RNA-Binding Proteins
  • Tubulin Modulators
  • 3-tritylthio-L-alanine
  • Vincristine
  • Kinesins
  • Cysteine
  • Paclitaxel
  • Cisplatin