2DLA

Primase large subunit amino terminal domain from Pyrococcus horikoshii

Summary for 2DLA

• Entry DOI: 10.2210/pdb2dla/pdb
• Descriptor: 397aa long hypothetical protein (2 entities in total)
• Functional Keywords: helix bundle, twisted beta-sheet, replication
• Biological source: Pyrococcus horikoshii
• Total number of polymer chains: 3
• Total formula weight: 78504.85

Authors

Ito, N. (deposition date: 2006-04-17, release date: 2007-02-27, Last modification date: 2024-04-03)

Primary citation

Ito, N.,Matsui, I.,Matsui, E.
Molecular basis for the subunit assembly of the primase from an archaeon Pyrococcus horikoshii
Febs J., 274:1340-1351, 2007

PubMed Abstract: Archaeal/eukaryotic primases form a heterodimer consisting of a small catalytic subunit (PriS) and a large subunit (PriL). The heterodimer complex synthesizes primer oligoribonucleotides that are required for chromosomal replication. Here, we describe crystallographic and biochemical studies of the N-terminal domain (NTD) of PriL (PriL(NTD); residues 1-222) that bind to PriS from a hyperthermophilic archaeon, Pyrococcus horikoshii, at 2.9 A resolution. The PriL(NTD) structure consists of two subdomains, the helix-bundle and twisted-strand domains. The latter is structurally flexible, and is expected to contain a PriS interaction site. Pull-down and surface plasmon resonance analyses of structure-based deletion and alanine scanning mutants showed that the conserved hydrophobic Tyr155-Tyr156-Ile157 region near the flexible region is the PriS-binding site, as the Y155A/Y156A/I157A mutation markedly reduces PriS binding, by 1000-fold. These findings and a structural comparison with a previously reported PriL(NTD)-PriS complex suggest that the presented alternative conformations of the twisted-strand domain facilitate the heterodimer assembly.

PubMed: 17286576
DOI: 10.1111/j.1742-4658.2007.05690.x

Experimental method

X-RAY DIFFRACTION (2.9 Å)