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Cdc48/p97-Ufd1-Npl4 antagonizes Aurora B during chromosome segregation in HeLa cells

Grzegorz Dobrynin1, Oliver Popp, Tina Romer

  • 1Centre for Medical Biotechnology, University of Duisburg-Essen, 45117 Essen, Germany.

Journal of Cell Science
|April 14, 2011
PubMed

Insights

The Cdc48/p97-Ufd1-Npl4 complex regulates Aurora B kinase activity on chromosomes during early mitosis. This regulation is vital for accurate chromosome segregation and preventing errors like missegregated chromosomes.

Area of Science:

  • Cell Biology
  • Molecular Biology
  • Genetics

Background:

  • The AAA+ ATPase Cdc48/p97 (VCP) and its adapter Ufd1-Npl4 are known to remove Aurora B kinase from chromatin during mitotic exit in Xenopus egg extracts.
  • Aurora B kinase plays a critical role in chromosome segregation during mitosis.

Purpose of the Study:

  • To investigate the role of the Ufd1-Npl4 complex in regulating Aurora B kinase activity on chromosomes during early mitosis in human somatic cells (HeLa cells).
  • To determine the impact of Ufd1-Npl4 on chromosome segregation fidelity.

Main Methods:

  • Depletion of Ufd1-Npl4 using small interfering RNA (siRNA) in HeLa cells.
  • Analysis of chromosome alignment, anaphase progression, and chromosome segregation.
  • Assessment of Aurora B levels and activity on chromosomes.
  • Use of the Aurora B inhibitor hesperadin to probe functional relationships.

Main Results:

  • Ufd1-Npl4 depletion caused chromosome alignment and anaphase defects, leading to missegregated chromosomes and multi-lobed nuclei.
  • Depletion resulted in increased Aurora B levels and activity on prometaphase and metaphase chromosomes.
  • Inhibition of Aurora B partially rescued chromosome alignment defects in Ufd1-Npl4-depleted cells, and vice versa.

Conclusions:

  • The Cdc48/p97-Ufd1-Npl4 complex acts as a crucial negative regulator of Aurora B kinase activity on chromosomes early in mitosis in human cells.
  • Restraining Aurora B activity on chromosomes is essential for ensuring faithful chromosome segregation.

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