Nucleic Acids Research Advance Access originally published online on November 10, 2006
Nucleic Acids Research 2006 34(21):6298-6304; doi:10.1093/nar/gkl890
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Nucleic Acids Research, 2006, Vol. 34, No. 21 6298-6304
© 2006 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.
Molecular Biology |
DAXX interacts with phage 
C31 integrase and inhibits recombination
1 State Key Laboratory of Genetic Engineering, Institute of Genetics, School of Life Sciences, Fudan University China 2 Department of Surgery, University of British Columbia Vancouver, Canada
*To whom correspondence should be addressed. Tel/Fax: +86 21 65649899; Email: Jlxue{at}fudan.ac.cn
Received July 10, 2006. Revised October 8, 2006. Accepted October 9, 2006.
Phage 
C31 integrase has potential as a means of inserting therapeutic genes into specific sites in the human genome. However, the possible interactions between C31 integrase and cellular proteins have never been investigated. Using pLexA-C31 integrase as bait, we screened a pB42AD-human fetal brain cDNA library for potential interacting cellular proteins. Among 61 positives isolated from 106 independent clones, 51 contained DAXX C-terminal fragments. The strong interaction between DAXX and C31 was further confirmed by co-immunoprecipitation. Deletion analysis revealed that the fas-binding domain of DAXX is also the region for C31 binding. Hybridization between a C31 integrase peptide array and an HEK293 cell extract revealed that a tetramer, 451RFGK454, in the C-terminus of C31 is responsible for the interaction with DAXX. This tetramer is also necessary for C31 integrase activity as removal of this tetramer resulted in a complete loss of integrase activity. Co-expression of DAXX with C31 integrase in a HEK293-derived C31 integrase activity reporter cell line significantly reduced the C31-mediated recombination rate. Knocking down DAXX with a DAXX-specific duplex RNA resulted in increased recombination efficiency. Therefore, endogenous DAXX may interact with C31 causing a mild inhibition in the integration efficiency. This is the first time that C31 was shown to interact with an important cellular protein and the potential effect of this interaction should be further studied.
*Correspondence may also be addressed to William Jia. Tel: +604 822 0728; Fax: +604 322 0640; Email: wjia{at}interchange.ubc.ca
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