Microtubule interfering agents and KSP inhibitors induce the phosphorylation of the nuclear protein p54(nrb), an event linked to G2/M arrest.
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).
Casado
Pedro
P
Departamento de Bioquímica y Biología Molecular, Instituto Universitario de Oncología del Principado de Asturias, Universidad de Oviedo, 33071, Oviedo, Spain.
Prado
Miguel A
MA
Zuazua-Villar
Pedro
P
Del Valle
Eva
E
Artime
Noelia
N
Cabal-Hierro
Lucía
L
Rupérez
Patricia
P
Burlingame
Alma L
AL
Lazo
Pedro S
PS
Ramos
Sofía
S
D000972:Antineoplastic Agents, Phytogenic / Q000494:pharmacology*; D002460:Cell Line; D002945:Cisplatin / Q000494:pharmacology; D003545:Cysteine / Q000031:analogs & derivatives / Q000494:pharmacology; D004268:DNA-Binding Proteins; D015180:Electrophoresis, Gel, Two-Dimensional; D016195:G2 Phase / Q000187:drug effects / Q000502:physiology*; D006801:Humans; D016547:Kinesins / Q000037:antagonists & inhibitors / Q000378:metabolism*; D008870:Microtubules; D008938:Mitosis / Q000502:physiology*; D050258:Mitosis Modulators / Q000494:pharmacology; D034521:Nuclear Matrix-Associated Proteins / Q000378:metabolism*; D050810:Octamer Transcription Factors / Q000378:metabolism*; D017239:Paclitaxel / Q000494:pharmacology; D010766:Phosphorylation; D016601:RNA-Binding Proteins / Q000378:metabolism*; D053719:Tandem Mass Spectrometry; D050257:Tubulin Modulators / Q000494:pharmacology*; D014750:Vincristine / Q000494:pharmacology
D000972:Antineoplastic Agents, Phytogenic; D004268:DNA-Binding Proteins; C470243:KIF11 protein, human; D050258:Mitosis Modulators; C083214:NONO protein, human; D034521:Nuclear Matrix-Associated Proteins; D050810:Octamer Transcription Factors; D016601:RNA-Binding Proteins; D050257:Tubulin Modulators; C003724:3-tritylthio-L-alanine; D014750:Vincristine; D016547:Kinesins; D003545:Cysteine; D017239:Paclitaxel; D002945:Cisplatin