TPEN, a Zn2+/Fe2+ chelator with low affinity for Ca2+, inhibits lamin assembly, destabilizes nuclear architecture and may independently protect nuclei from apoptosis in vitro

D. K. Shumaker, L. R. Vann, M. W. Goldberg, T. D. Allen, Katherine Lee Wilson

Research output: Contribution to journalArticle

Abstract

We used Xenopus egg extracts to examine the effects of TPEN, a chelator with strong affinities for Zn2+, Fe2+, and Mn2+ on nuclear assembly in vitro. At concentrations above 1 mM, TPEN blocked the assembly of the nuclear lamina and produced nuclei that were profoundly sensitive to stress-induced balloon-like 'shedding' of nuclear membranes away from chromatin-associated membranes. TPEN-arrested nuclei were also defective for DNA replication, which could be explained as secondary to the lack of a lamina. Imaging of TPEN-arrested nuclei by field emission in-lens scanning electron microscopy (FEISEM) revealed clustered, structurally-perturbed nuclear pore complexes. TPEN-arrested nuclei were defective in the accumulation of fluorescent karyophilic proteins. All detectable effects caused by TPEN were downstream of the effects of BAPTA, a Ca2+/Zn2+ chelator that blocks pore complex assembly at two distinct early stages. Surprisingly, TPEN-arrested nuclei, but not control nuclei, remained active for replication in apoptotic extracts, as assayed by [32P]-dCTP incorporation into high molecular weight DNA, suggesting that TPEN blocks a metal-binding protein(s) required for nuclear destruction during programmed cell death.

Original languageEnglish (US)
Pages (from-to)151-164
Number of pages14
JournalCell Calcium
Volume23
Issue number2-3
DOIs
StatePublished - 1998

Fingerprint

Lamins
Chelating Agents
Apoptosis
Nuclear Lamina
Nuclear Pore
N,N,N',N'-tetrakis(2-pyridylmethyl)ethylenediamine
In Vitro Techniques
Nuclear Envelope
Xenopus
DNA Replication
Electron Scanning Microscopy
Lenses
Chromatin
Ovum
Carrier Proteins
Cell Death
Molecular Weight
Metals
Membranes
DNA

ASJC Scopus subject areas

  • Cell Biology
  • Endocrinology

Cite this

TPEN, a Zn2+/Fe2+ chelator with low affinity for Ca2+, inhibits lamin assembly, destabilizes nuclear architecture and may independently protect nuclei from apoptosis in vitro. / Shumaker, D. K.; Vann, L. R.; Goldberg, M. W.; Allen, T. D.; Wilson, Katherine Lee.

In: Cell Calcium, Vol. 23, No. 2-3, 1998, p. 151-164.

Research output: Contribution to journalArticle

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