Identification and autonomous replication capability of a chromosomal replication origin from the archaeon Sulfolobus solfataricus

Patrizia Contursi, Francesca M. Pisani, Andrei Grigoriev, Raffaele Cannio, Simonetta Bartolucci, Mosè Rossi

Research output: Contribution to journalArticlepeer-review

21 Scopus citations

Abstract

Here, we describe the identification of a chromosomal DNA replication origin (oriC) from the hyperthermophilic archaeon Sulfolobus solfataricus (subdomain of Crenarchaeota). By means of a cumulative GC-skew analysis of the Sulfolobus genome sequence, a candidate oriC was mapped within a 1.12-kb region located between the two divergently transcribed MCM- and cdc6-like genes. We demonstrated that plasmids containing the Sulfolobus oriC sequence and a hygromycin-resistance selectable marker were maintained in an episomal state in transformed S. solfataricus cells under selective pressure. The proposed location of the origin was confirmed by 2-D gel electrophoresis experiments. This is the first report on the functional cloning of a chromosomal oriC from an archaeon and represents an important step toward the reconstitution of an archaeal in vitro DNA replication system.

Original languageEnglish (US)
Pages (from-to)385-391
Number of pages7
JournalExtremophiles
Volume8
Issue number5
DOIs
StatePublished - Oct 2004
Externally publishedYes

All Science Journal Classification (ASJC) codes

  • Microbiology
  • Molecular Medicine

Keywords

  • Archaea
  • DNA replication
  • GC-skew analysis
  • Replication origin
  • Sulfolobus solfataricus

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