Identification and autonomous replication capability of a chromosomal replication origin from the archaeon Sulfolobus solfataricus.
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.
Contursi
Patrizia
P
Dipartimento di Chimica Biologica, Università degli Studi di Napoli, Via Mezzocannone, 16, 80134, Napoli, Italy.
Pisani
Francesca M
FM
Grigoriev
Andrei
A
Cannio
Raffaele
R
Bartolucci
Simonetta
S
Rossi
Mosè
M
D001105:Archaea / Q000235:genetics; D002453:Cell Cycle; D019847:Chromosomes, Archaeal / Q000235:genetics; D003001:Cloning, Molecular; D004247:DNA / Q000235:genetics; D004261:DNA Replication; D019641:DNA, Archaeal / Q000235:genetics*; D015180:Electrophoresis, Gel, Two-Dimensional; D005822:Genetic Vectors; D008957:Models, Genetic; D010957:Plasmids / Q000378:metabolism; D011312:Pressure; D018741:Replication Origin; D048229:Sulfolobus solfataricus / Q000235:genetics*
D019641:DNA, Archaeal; D004247:DNA