5UAE

Crystal structure of the coiled-coil domain from Listeria Innocua Phage Integrase (Trigonal Form)

Summary for 5UAE

• Entry DOI: 10.2210/pdb5uae/pdb
• Related: 5U96
• Descriptor: Putative integrase, CITRATE ANION (3 entities in total)
• Functional Keywords: site-specific recombination, coiled-coil, recombination
• Biological source: Listeria innocua
• Total number of polymer chains: 4
• Total formula weight: 30402.25

Authors

Gupta, K.,Sharp, R.,Yuan, J.B.,Van Duyne, G.D. (deposition date: 2016-12-19, release date: 2017-05-24, Last modification date: 2023-10-04)

Primary citation

Gupta, K.,Sharp, R.,Yuan, J.B.,Li, H.,Van Duyne, G.D.
Coiled-coil interactions mediate serine integrase directionality.
Nucleic Acids Res., 45:7339-7353, 2017

PubMed Abstract: Serine integrases are bacteriophage enzymes that carry out site-specific integration and excision of their viral genomes. The integration reaction is highly directional; recombination between the phage attachment site attP and the host attachment site attB to form the hybrid sites attL and attR is essentially irreversible. In a recent model, extended coiled-coil (CC) domains in the integrase subunits are proposed to interact in a way that favors the attPxattB reaction but inhibits the attLxattR reaction. Here, we show for the Listeria innocua integrase (LI Int) system that the CC domain promotes self-interaction in isolated Int and when Int is bound to attachment sites. Three independent crystal structures of the CC domain reveal the molecular nature of the CC dimer interface. Alanine substitutions of key residues in the interface support the functional significance of the structural model and indicate that the same interaction is responsible for promoting integration and for inhibiting excision. An updated model of a LI Int•attL complex that incorporates the high resolution CC dimer structure provides insights that help to explain the unusual CC dimer structure and potential sources of stability in Int•attL and Int•attR complexes. Together, the data provide a molecular basis for understanding serine integrase directionality.

PubMed: 28549184
DOI: 10.1093/nar/gkx474

Experimental method

X-RAY DIFFRACTION (2.75 Å)