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J. Biol. Chem., Vol. 277, Issue 12, 10209-10219, March 22, 2002
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From the Laboratoire de Pharmacologie et de Biologie
Structurale, CNRS UMR 5089, 205 route de Narbonne, 31077 Toulouse
Cedex, France and Nucleolin is one of the most abundant
non-ribosomal proteins of the nucleolus. Several studies in
vitro have shown that nucleolin is involved in several steps of
ribosome biogenesis, including the regulation of rDNA transcription,
rRNA processing, and ribosome assembly. However, the different steps of
ribosome biogenesis are highly coordinated, and therefore it is not
clear to what extent nucleolin is involved in each of these steps. It
has been proposed that the interaction of nucleolin with the rDNA
sequence and with nascent pre-rRNA leads to the blocking of RNA
polymerase I (RNA pol I) transcription. To test this model and to get
molecular insights into the role of nucleolin in RNA pol I
transcription, we studied the function of nucleolin in
Xenopus oocytes. We show that injection of a 2-4-fold
excess of Xenopus or hamster nucleolin in stage VI
Xenopus oocytes reduces the accumulation of 40 S pre-rRNA 3-fold, whereas transcription by RNA polymerase II and III is not affected. Direct analysis of rDNA transcription units by electron microscopy reveals that the number of polymerase complexes/rDNA unit is
drastically reduced in the presence of increased amounts of nucleolin
and corresponds to the level of reduction of 40 S pre-rRNA.
Transcription from DNA templates containing various combinations of RNA
polymerase I or II promoters in fusion with rDNA or CAT sequences was
analyzed in the presence of elevated amounts of nucleolin. It was shown
that nucleolin leads to transcription repression from a minimal
polymerase I promoter, independently of the nature of the RNA sequence
that is transcribed. Therefore, we propose that nucleolin affects RNA
pol I transcription by acting directly on the transcription machinery
or on the rDNA promoter sequences and not, as previously thought,
through interaction with the nascent pre-rRNA.
Repression of RNA Polymerase I Transcription by Nucleolin Is
Independent of the RNA Sequence That Is Transcribed*
§
Laboratoire de Biologie
Moléculaire et Cellulaire, Ecole Normale Supérieure de
Lyon, CNRS UMR 5665, 46 allée d'Italie,
69364 Lyon Cedex, France
*
This work was supported by grants from the Région
Midi-Pyrénées, University Paul Sabatier (Toulouse), the
CNRS (Action Thématique Incitative sun Programme et Equipe
(ATIPE)), and the Association pour la Recherche contre le
Cancer.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 © 2002 by The American Society for Biochemistry and Molecular Biology, Inc.
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