Selection of homeotic proteins for binding to a human DNA replication origin

Elisa De Stanchina, Davide Gabellini, Paolo Norio, Mauro Giacca, Fiorenzo A. Peverali, Silvano Riva, Arturo Falaschi, Giuseppe Biamonti

Research output: Contribution to journalArticlepeer-review

Abstract

We have previously shown that a cell cycle-dependent nucleoprotein complex assembles in vivo on a 74 bp sequence within the human DNA replication origin associated to the Lamin B2 gene. Here, we report the identification, using a one-hybrid screen in yeast, of three proteins interacting with the 74 bp sequence. All of them, namely HOXA13, HOXC10 and HOXC13, are orthologues of the Abdominal-B gene of Drosophila melanogaster and are members of the homeogene family of developmental regulators. We describe the complete open reading frame sequence of HOXC10 and HOXC13 along with the structure of the HoxC13 gene. The specificity of binding of these two proteins to the Lamin B2 origin is confirmed by both band-shift and in vitro footprinting assays. In addition, the ability of HOXC10 and HOXC13 to increase the activity of a promoter containing the 74 bp sequence, as assayed by CAT-assay experiments, demonstrates a direct interaction of these homeoproteins with the origin sequence in mammalian cells. We also show that HOXC10 expression is cell-type-dependent and positively correlates with cell proliferation. (C) 2000 Academic Press.

Original languageEnglish
Pages (from-to)667-680
Number of pages14
JournalJournal of Molecular Biology
Volume299
Issue number3
DOIs
Publication statusPublished - Jun 9 2000

Keywords

  • DNA replication origin
  • Homeoproteins
  • Lamin B2
  • One-hybrid

ASJC Scopus subject areas

  • Virology

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