PDBsum entry 2gdj

Recombination

PDB id: 2gdj

Contents

Protein chain

237 a.a. *

Ligands

ANP

Metals

_MG ×2

* Residue conservation analysis

PDB id:

2gdj

Name:

Recombination

Title:

Delta-62 rada recombinase in complex with amp-pnp and magnesium

Structure:

DNA repair and recombination protein rada. Chain: a. Fragment: atpase domain, residues 62-322. Engineered: yes

Source:

Methanococcus voltae. Organism_taxid: 2188. Gene: rada. Expressed in: escherichia coli. Expression_system_taxid: 562.

Resolution:

2.50Å R-factor: 0.210 R-free: 0.268

Authors:

Y.Wu,X.Qian,Y.He,Y.Luo

Key ref:

V.E.Galkin et al. (2006). The Rad51/RadA N-terminal domain activates nucleoprotein filament ATPase activity. Structure, 14, 983-992. PubMed id: 16765891 DOI: 10.1016/j.str.2006.04.001

Date:

16-Mar-06 Release date: 16-May-06

Protein chain

O73948  (RADA_METVO) -  DNA repair and recombination protein RadA from Methanococcus voltae

Seq: 322 a.a.

Struc: 237 a.a.

DOI no: 10.1016/j.str.2006.04.001

Structure 14:983-992 (2006)

PubMed id: 16765891

The Rad51/RadA N-terminal domain activates nucleoprotein filament ATPase activity.

V.E.Galkin, Y.Wu, X.P.Zhang, X.Qian, Y.He, X.Yu, W.D.Heyer, Y.Luo, E.H.Egelman.

ABSTRACT

Proteins in the RecA/RadA/Rad51 family form helical filaments on DNA that function in homologous recombination. While these proteins all have the same highly conserved ATP binding core, the RadA/Rad51 proteins have an N-terminal domain that shows no homology with the C-terminal domain found in RecA. Both the Rad51 N-terminal and RecA C-terminal domains have been shown to bind DNA, but no role for these domains has been established. We show that RadA filaments can be trapped in either an inactive or active conformation with respect to the ATPase and that activation involves a large rotation of the subunit aided by the N-terminal domain. The G103E mutation within the yeast Rad51 N-terminal domain inactivates the filament by failing to make proper contacts between the N-terminal domain and the core. These results show that the N-terminal domains play a regulatory role in filament activation and highlight the modular architecture of the recombination proteins.

Selected figure(s)

Figure 1.
Figure 1. Crystal Structure of the RadA-Δ62 Fragment
(A) The RadA-Δ62 conformation (cyan) is essentially identical to the structure of the corresponding core in the full-length protein (gold), except for the polymerization motif around Phe-64 and a short helical hinge.
(B) The ATPase site of the RadA-Δ62 filament resembles that of the previously determined full-length RadA structure. The AMP-PNP (in cyan) is found between two adjacent protomers. The yellow protomer contributes the triphosphate-wrapping P loop, base-stacking Arg-158, and the catalytic Glu-151 and Gln-257. The gray protomer contacts the ATP analog through the ATP cap (residues Asp-302 to Asp-308) and the C-terminal elbow of the L2 region (residues His-280 to Arg-285). The magnesium ion is shown in red.

Figure 8.
Figure 8. ATPase Activity of Wild-Type and G103E Mutant Yeast Rad51 Protein
The initial activity seen with the G103E mutant, followed by a plateau showing no further activity, is consistent with a single hydrolytic event with no subsequent turnover.

The above figures are reprinted by permission from Cell Press: Structure (2006, 14, 983-992) copyright 2006.

Figures were selected by an automated process.