Nucleic Acids Research Advance Access originally published online on May 23, 2008
Nucleic Acids Research 2008 36(12):3879-3891; doi:10.1093/nar/gkn269
| ||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Nucleic Acids Research, 2008, Vol. 36, No. 12 3879-3891
© 2008 The Author(s)
This is an Open Access article distributed under the terms of the Creative Commons Attribution Non-Commercial License (http://creativecommons.org/licenses/by-nc/2.0/uk/) which permits unrestricted non-commercial use, distribution, and reproduction in any medium, provided the original work is properly cited.
Nucleic Acid Enzymes |
A motif in the C-terminal domain of 
C31 integrase controls the directionality of recombination
1Institute of Medical Sciences, University of Aberdeen, Foresterhill, Aberdeen AB252ZD and 2Institute of Genetics, University of Nottingham, Nottingham NG7 2UH, UK
*To whom correspondence should be addressed. Tel: +44 1224 555739; Fax: +44 1224 555844; Email: maggie.smith{at}abdn.ac.uk
Received March 1, 2008. Revised March 12, 2008. Accepted April 22, 2008.
Bacteriophage 
C31 encodes an integrase, which acts on the phage and host attachment sites, attP and attB, to form an integrated prophage flanked by attL and attR. In the absence of accessory factors, C31 integrase cannot catalyse attL x attR recombination to excise the prophage. To understand the mechanism of directionality, mutant integrases were characterized that were active in excision. A hyperactive integrase, Int E449K, gained the ability to catalyse attL x attR, attL x attL and attR x attR recombination whilst retaining the ability to recombine attP x attB. A catalytically defective derivative of this mutant, Int S12A, E449K, could form stable complexes with attP/attB, attL/attR, attL/attL and attR/attR under conditions where Int S12A only complexed with attP/attB. Further analysis of the Int E449K-attL/attR synaptic events revealed a preference for one of the two predicted synapse structures with different orientations of the attL/attR sites. Several amino acid substitutions conferring hyperactivity, including E449K, were localized to one face of a predicted coiled-coil motif in the C-terminal domain. This work shows that a motif in the C-terminal domain of C31 integrase controls the formation of the synaptic interface in both integration and excision, possibly through a direct role in protein–protein interactions.
CiteULike 
Connotea 
Del.icio.us What's this?
This article has been cited by other articles:
|
|
 |
|
  A. R. McEwan, P. A. Rowley, and M. C. M. Smith DNA binding and synapsis by the large C-terminal domain of {phi}C31 integrase Nucleic Acids Res., August 1, 2009; 37(14): 4764 - 4773. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
F. J. Olorunniji, J. He, S. V.C.T. Wenwieser, M. R. Boocock, and W. M. Stark Synapsis and catalysis by activated Tn3 resolvase mutants Nucleic Acids Res., December 1, 2008; 36(22): 7181 - 7191. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 P. A. Rowley and M. C. M. Smith Role of the N-Terminal Domain of {phi}C31 Integrase in attB-attP Synapsis J. Bacteriol., October 15, 2008; 190(20): 6918 - 6921. [Abstract] [Full Text] [PDF]
|
|