shared active site architecture between the large subunit of eukaryotic primase and dna photolyase大亚基之间的共享活动网站架构真核引发酶和dna光裂合酶.pdfVIP

Shared Active Site Architecture between the Large Subunit of Eukaryotic Primase and DNA Photolyase ¤ Ludovic Sauguet, Sebastian Klinge , Rajika L. Perera, Joseph D. Maman, Luca Pellegrini* Department of Biochemistry, University of Cambridge, Cambridge, United Kingdom Abstract Background: DNA synthesis during replication relies on RNA primers synthesised by the primase, a specialised DNA- dependent RNA polymerase that can initiate nucleic acid synthesis de novo. In archaeal and eukaryotic organisms, the primase is a heterodimeric enzyme resulting from the constitutive association of a small (PriS) and large (PriL) subunit. The ability of the primase to initiate synthesis of an RNA primer depends on a conserved Fe-S domain at the C-terminus of PriL (PriL-CTD). However, the critical role of the PriL-CTD in the catalytic mechanism of initiation is not understood. ˚ Methodology/Principal Findings: Here we report the crystal structure of the yeast PriL-CTD at 1.55 A resolution. The structure reveals that the PriL-CTD folds in two largely independent alpha-helical domains joined at their interface by a [4Fe- 4S] cluster. The larger N-terminal domain represents the most conserved portion of the PriL-CTD, whereas the smaller C- terminal domain is largely absent in archaeal PriL. Unexpectedly, the N-terminal domain reveals a striking structural similarity with the active site region of the DNA photolyase/cryptochrome family of flavoproteins. The region of similarity includes PriL-CTD residues that are known to be essential for initiation of RNA primer synthesis by the primase. Conclusion/Significance: Our study rep