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Title: Structural Basis for Catalytic Activation of a Serine Recombinase

Abstract

Sin resolvase is a site-specific serine recombinase that is normally controlled by a complex regulatory mechanism. A single mutation, Q115R, allows the enzyme to bypass the entire regulatory apparatus, such that no accessory proteins or DNA sites are required. Here, we present a 1.86 {angstrom} crystal structure of the Sin Q115R catalytic domain, in a tetrameric arrangement stabilized by an interaction between Arg115 residues on neighboring subunits. The subunits have undergone significant conformational changes from the inactive dimeric state previously reported. The structure provides a new high-resolution view of a serine recombinase active site that is apparently fully assembled, suggesting roles for the conserved active site residues. The structure also suggests how the dimer-tetramer transition is coupled to assembly of the active site. The tetramer is captured in a different rotational substate than that seen in previous hyperactive serine recombinase structures, and unbroken crossover site DNA can be readily modeled into its active sites.

Authors:
; ; ; ;  [1];  [2]
  1. (Glasgow)
  2. (
Publication Date:
Research Org.:
Argonne National Lab. (ANL), Argonne, IL (United States). Advanced Photon Source (APS)
Sponsoring Org.:
OTHERNIH
OSTI Identifier:
1027653
Resource Type:
Journal Article
Journal Name:
Structure
Additional Journal Information:
Journal Volume: 19; Journal Issue: (6) ; 06, 2011
Country of Publication:
United States
Language:
ENGLISH
Subject:
59 BASIC BIOLOGICAL SCIENCES; 60 APPLIED LIFE SCIENCES; CONFORMATIONAL CHANGES; CRYSTAL STRUCTURE; DNA; ENZYMES; PROTEINS; RESIDUES; SERINE

Citation Formats

Keenholtz, Ross A., Rowland, Sally-J., Boocock, Martin R., Stark, W. Marshall, Rice, Phoebe A., and UC). Structural Basis for Catalytic Activation of a Serine Recombinase. United States: N. p., 2014. Web. doi:10.1016/j.str.2011.03.017.
Keenholtz, Ross A., Rowland, Sally-J., Boocock, Martin R., Stark, W. Marshall, Rice, Phoebe A., & UC). Structural Basis for Catalytic Activation of a Serine Recombinase. United States. doi:10.1016/j.str.2011.03.017.
Keenholtz, Ross A., Rowland, Sally-J., Boocock, Martin R., Stark, W. Marshall, Rice, Phoebe A., and UC). Thu . "Structural Basis for Catalytic Activation of a Serine Recombinase". United States. doi:10.1016/j.str.2011.03.017.
@article{osti_1027653,
title = {Structural Basis for Catalytic Activation of a Serine Recombinase},
author = {Keenholtz, Ross A. and Rowland, Sally-J. and Boocock, Martin R. and Stark, W. Marshall and Rice, Phoebe A. and UC)},
abstractNote = {Sin resolvase is a site-specific serine recombinase that is normally controlled by a complex regulatory mechanism. A single mutation, Q115R, allows the enzyme to bypass the entire regulatory apparatus, such that no accessory proteins or DNA sites are required. Here, we present a 1.86 {angstrom} crystal structure of the Sin Q115R catalytic domain, in a tetrameric arrangement stabilized by an interaction between Arg115 residues on neighboring subunits. The subunits have undergone significant conformational changes from the inactive dimeric state previously reported. The structure provides a new high-resolution view of a serine recombinase active site that is apparently fully assembled, suggesting roles for the conserved active site residues. The structure also suggests how the dimer-tetramer transition is coupled to assembly of the active site. The tetramer is captured in a different rotational substate than that seen in previous hyperactive serine recombinase structures, and unbroken crossover site DNA can be readily modeled into its active sites.},
doi = {10.1016/j.str.2011.03.017},
journal = {Structure},
number = (6) ; 06, 2011,
volume = 19,
place = {United States},
year = {2014},
month = {10}
}