2FAO

Crystal Structure of Pseudomonas aeruginosa LigD polymerase domain

Summary for 2FAO

• Entry DOI: 10.2210/pdb2fao/pdb
• Related: 2FAQ 2FAR
• Descriptor: probable ATP-dependent DNA ligase, SULFATE ION (3 entities in total)
• Functional Keywords: polymerase, primase, ligase, nhej, hydrolase-transferase complex, hydrolase/transferase
• Biological source: Pseudomonas aeruginosa
• Total number of polymer chains: 2
• Total formula weight: 68866.63

Authors

Zhu, H.,Nandakumar, J.,Aniukwu, J.,Wang, L.K.,Glickman, M.S.,Lima, C.D.,Shuman, S. (deposition date: 2005-12-07, release date: 2006-05-23, Last modification date: 2024-02-14)

Primary citation

Zhu, H.,Nandakumar, J.,Aniukwu, J.,Wang, L.K.,Glickman, M.S.,Lima, C.D.,Shuman, S.
Atomic structure and nonhomologous end-joining function of the polymerase component of bacterial DNA ligase D
Proc.Natl.Acad.Sci.USA, 103:1711-1716, 2006

PubMed Abstract: DNA ligase D (LigD) is a large polyfunctional protein that participates in a recently discovered pathway of nonhomologous end-joining in bacteria. LigD consists of an ATP-dependent ligase domain fused to a polymerase domain (Pol) and a phosphoesterase module. The Pol activity is remarkable for its dependence on manganese, its ability to perform templated and nontemplated primer extension reactions, and its preference for adding ribonucleotides to blunt DNA ends. Here we report the 1.5-A crystal structure of the Pol domain of Pseudomonas LigD and its complexes with manganese and ATP/dATP substrates, which reveal a minimized polymerase with a two-metal mechanism and a fold similar to that of archaeal DNA primase. Mutational analysis highlights the functionally relevant atomic contacts in the active site. Although distinct nucleoside conformations and contacts for ATP versus dATP are observed in the cocrystals, the functional analysis suggests that the ATP-binding mode is the productive conformation for dNMP and rNMP incorporation. We find that a mutation of Mycobacterium LigD that uniquely ablates the polymerase activity results in increased fidelity of blunt-end double-strand break repair in vivo by virtue of eliminating nucleotide insertions at the recombination junctions. Thus, LigD Pol is a direct catalyst of mutagenic nonhomologous end-joining in vivo. Our studies underscore a previously uncharacterized role for the primase-like polymerase family in DNA repair.

PubMed: 16446439
DOI: 10.1073/pnas.0509083103

Experimental method

X-RAY DIFFRACTION (1.5 Å)