3M1M

Crystal structure of the primase-polymerase from Sulfolobus islandicus

Summary for 3M1M

• Entry DOI: 10.2210/pdb3m1m/pdb
• Descriptor: ORF904, ZINC ION, GLYCEROL, ... (5 entities in total)
• Functional Keywords: primase, polymerase, replication
• Biological source: Sulfolobus islandicus
• Total number of polymer chains: 1
• Total formula weight: 39221.34

Authors

Vannini, A.,Beck, K.,Lipps, G.,Cramer, P. (deposition date: 2010-03-05, release date: 2010-06-16, Last modification date: 2024-10-30)

Primary citation

Beck, K.,Vannini, A.,Cramer, P.,Lipps, G.
The archaeo-eukaryotic primase of plasmid pRN1 requires a helix bundle domain for faithful primer synthesis
Nucleic Acids Res., 38:6707-6718, 2010

PubMed Abstract: The plasmid pRN1 encodes for a multifunctional replication protein with primase, DNA polymerase and helicase activity. The minimal region required for primase activity encompasses amino-acid residues 40-370. While the N-terminal part of that minimal region (residues 47-247) folds into the prim/pol domain and bears the active site, the structure and function of the C-terminal part (residues 248-370) is unknown. Here we show that the C-terminal part of the minimal region folds into a compact domain with six helices and is stabilized by a disulfide bond. Three helices superimpose well with the C-terminal domain of the primase of the bacterial broad host range plasmid RSF1010. Structure-based site-directed mutagenesis shows that the C-terminal helix of the helix bundle domain is required for primase activity although it is distant to the active site in the crystallized conformation. Furthermore, we identified mutants of the C-terminal domain, which are defective in template binding, dinucleotide formation and conformation change prior to DNA extension.

PubMed: 20511586
DOI: 10.1093/nar/gkq447

Experimental method

X-RAY DIFFRACTION (1.85 Å)