The GINS complex from Pyrococcus furiosus stimulates the MCM helicase activity.
Pyrococcus furiosus, a hyperthermophilic Archaea, has homologs of the eukaryotic MCM (mini-chromosome maintenance) helicase and GINS complex. The MCM and GINS proteins are both essential factors to initiate DNA replication in eukaryotic cells. Many biochemical characterizations of the replication-related proteins have been reported, but it has not been proved that the homologs of each protein are also essential for replication in archaeal cells. Here, we demonstrated that the P. furiosus GINS complex interacts with P. furiosus MCM. A chromatin immunoprecipitation assay revealed that the GINS complex is detected preferentially at the oriC region on Pyrococcus chromosomal DNA during the exponential growth phase but not in the stationary phase. Furthermore, the GINS complex stimulates both the ATPase and DNA helicase activities of MCM in vitro. These results strongly suggest that the archaeal GINS is involved in both the initiation and elongation processes of DNA replication in P. furiosus, as observed in eukaryotic cells.
Yoshimochi
Takehiro
T
Department of Genetic Resources Technology, Faculty of Agriculture, Kyushu University, 6-10-1 Fukuoka 812-8581, Japan.
Fujikane
Ryosuke
R
Department of Genetic Resources Technology, Faculty of Agriculture, Kyushu University, 6-10-1 Fukuoka 812-8581, Japan.
Kawanami
Miyuki
M
Department of Genetic Resources Technology, Faculty of Agriculture, Kyushu University, 6-10-1 Fukuoka 812-8581, Japan.
Matsunaga
Fujihiko
F
Department of Genetic Resources Technology, Faculty of Agriculture, Kyushu University, 6-10-1 Fukuoka 812-8581, Japan.
Ishino
Yoshizumi
Y
Department of Genetic Resources Technology, Faculty of Agriculture, Kyushu University, 6-10-1 Fukuoka 812-8581, Japan; BIRD-Japan Science and Technology Agency, 6-10-1 Fukuoka 812-8581, Japan. Electronic address: ishino@agr.kyushu-u.ac.jp.
D000251:Adenosine Triphosphatases / Q000378:metabolism; D019843:Archaeal Proteins / Q000737:chemistry / Q000302:isolation & purification / Q000378:metabolism*; D002868:Chromosomal Proteins, Non-Histone / Q000302:isolation & purification / Q000378:metabolism*; D017124:Conserved Sequence; D004247:DNA / Q000378:metabolism; D004265:DNA Helicases / Q000737:chemistry / Q000302:isolation & purification / Q000378:metabolism*; D019766:Genes, Archaeal; D047468:Immunoprecipitation; D008954:Models, Biological; D051738:Origin Recognition Complex; D011485:Protein Binding; D020836:Protein Structure, Quaternary; D020027:Pyrococcus furiosus / Q000201:enzymology* / Q000235:genetics / Q000254:growth & development; D012689:Sequence Homology, Nucleic Acid; D012996:Solutions; D020798:Two-Hybrid System Techniques
D019843:Archaeal Proteins; D002868:Chromosomal Proteins, Non-Histone; C114650:OriC chromosomal replication origin; D051738:Origin Recognition Complex; D012996:Solutions; D004247:DNA; D000251:Adenosine Triphosphatases; D004265:DNA Helicases