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.
Dipartimento di Chimica Biologica, Università degli Studi di Napoli, Via Mezzocannone, 16, 80134, Napoli, Italy.
Patrizia
P
Contursi
Francesca M
FM
Pisani
Andrei
A
Grigoriev
Raffaele
R
Cannio
Simonetta
S
Bartolucci
Mosè
M
Rossi
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