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Crystal structure of the Pyrococcus horikoshii DNA primase‐UTP complex: implications for the mechanism of primer synthesis
Author(s) -
Ito Nobutoshi,
Nureki Osamu,
Shirouzu Mikako,
Yokoyama Shigeyuki,
Hanaoka Fumio
Publication year - 2003
Publication title -
genes to cells
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.912
H-Index - 115
eISSN - 1365-2443
pISSN - 1356-9597
DOI - 10.1111/j.1365-2443.2003.00693.x
Subject(s) - primase , biology , dna polymerase , primer (cosmetics) , dna replication , dna clamp , dna , biochemistry , replisome , dna polymerase ii , pyrococcus horikoshii , circular bacterial chromosome , enzyme , rna , chemistry , gene , reverse transcriptase , organic chemistry
Background: In chromosomal DNA replication, DNA primase initiates the synthesis of a dinucleotide on a single‐stranded template DNA, and elongates it to form a primer RNA for the replicative DNA polymerase. Although the apo‐structure of an archaeal primase has been reported, the mechanism of primer synthesis by the eukaryotic‐type primase still remains to be elucidated. Results: In this study, we present the crystal structure of the eukaryotic‐type DNA primase from the hyperthermophilic archaeon ( Pyrococcus horikoshii ) with the uridine 5′‐triphosphate (UTP). In the present primase‐UTP complex, the primase binds the triphosphate moiety of the UTP at the active site, which includes Asp95, Asp97, and Asp280, the essential residues for the nucleotidyl transfer reaction. Conclusion: The nucleotide binding geometry in this complex explains the previous biochemical analyses of the eukaryotic primase. Based on the complex structure, we constructed a model between the DNA primase and a primer/template DNA for the primer synthesis. This model facilitates the comprehension of the reported features of DNA primase.