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J. Biol. Chem., Vol. 276, Issue 8, 5769-5778, February 23, 2001
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From the Two isoforms of DNA topoisomerase II (topo II)
have been identified in mammalian cells. While topo II
Involvement of DNA Topoisomerase II
in Neuronal
Differentiation*
§,
, and Department of Molecular Biology, Institute
of Cellular and Molecular Biology, the ¶ Third Department of
Anatomy, Okayama University Medical School, Okayama 700-8558, Japan and
the Laboratory of Medical Mycology, Research Institute for
Disease Mechanism and Control, Nagoya University School of Medicine,
Nagoya 466-8550, Japan
is essential
for chromosome segregation in mitotic cells, in vivo
function of topo II
remains to be clarified. Here we demonstrate
that the nucleoplasmic topo II, highly expressed in differentiating
cerebellar neurons, is the catalytically competent entity operating
directly on chromatin DNA in vivo. When the cells reached
terminal differentiation, this in vivo activity decreased
to a negligible level with concomitant loss of the nucleoplasmic
enzyme. Effects of topo II-specific inhibitors were analyzed in a
primary culture of cerebellar granule neurons that can mimic the
in vivo situation. Only the isoform was expressed in
granule cells differentiating in vitro. ICRF-193, a
catalytic topo II inhibitor, suppressed the transcriptional induction
of amphiphysin I which is essential for mature neuronal activity. The
effect decreased significantly as the cells differentiate. Expression
profiling with a cDNA macroarray showed that 18% of detectable
transcripts were up-regulated during the differentiation and one-third
of them were susceptible to ICRF-193. The results suggest that topo
II is involved in an early stage of granule cell differentiation by
potentiating inducible neuronal genes to become transcribable probably
through alterations in higher order chromatin structure.
*
This work was supported by grants-in-aid for Scientific
Research from the Ministry of Education, Science, Sports and Culture of
Japan (project number 11239206).The costs of publication of this
article were defrayed in part by the
payment of page charges. The article
must therefore be hereby marked
"advertisement" in accordance with 18 U.S.C. Section
1734 solely to indicate this fact.
Copyright © 2001 by The American Society for Biochemistry and Molecular Biology, Inc.
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