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Sp100 interacts with phage ΦC31 integrase to inhibit its recombination activity

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Lin Y., Li Z.-H., Wang J.-J., Xu G.-L., Shen Q., Tian L., Xue J.-L., Chen J.-Z.

2011

vol. 58

issue 1

67-73

Phage ΦC31 integrase is a potential vector for the insertion of therapeutic genes into specific sites in the human genome. To understand the mechanism involved in ΦC31 integrase-mediated recombination, it is important to understand the interaction between the integrase and cellular proteins. Using a yeast two-hybrid system with pLexA-ΦC31 integrase as bait, we screened a pB42AD human fetal brain cDNA library for potential interacting cellular proteins. From the 106 independent clones that were screened, 11 potential interacting clones were isolated, of which one encoded C-terminal fragment of Sp100. The interaction between Sp100 and ΦC31 integrase was further confirmed by yeast mating and co-immunoprecipitation assays. The hybridization between a ΦC31 integrase peptide array and an HEK293 cell extract revealed that residues 81RILN84 in the N-terminus of ΦC31 integrase are responsible for the interaction with Sp100. Knocking down endogenous Sp100 with Sp100-specific siRNA increased ΦC31 integrase-mediated recombination but did not impact reporter gene expression. Therefore, endogenous Sp100 may interact with ΦC31 integrase and inhibit the efficiency of ΦC31 integrase-mediated recombination.

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1 Acta Biochimica Polonica

1 2011

1 Chen J.-Z.

1 Li Z.-H.

1 Lin Y.

1 Shen Q.

1 Tian L.

1 Wang J.-J.

1 Xu G.-L.

1 Xue J.-L.

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Sp100 interacts with phage ΦC31 integrase to inhibit its recombination activity