Sp100 interacts with phage ΦC31 integrase to inhibit its recombination activity

• Autorzy: Lin Y. , Li Z. , Wang J.J. , Xu G.I. , Shen Q. , Tian L. , Xue J.I. , Chen J.
• Języki publikacji: EN

Abstrakty

EN

 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.

Słowa kluczowe

EN

cellular protein; gene; genome; human genome; integrase system; interaction; recombination; therapeutic gene; green fluorescent protein; red fluorescent protein; phenylmethylsulphonyl fluoride

• Wydawca: -
• Czasopismo: Acta Biochimica Polonica
• Rocznik: 2011
• Tom: 58
• Numer: 1
• Opis fizyczny: p.67-73,fig.,ref.

Twórcy

autor

Lin Y.

• State Key Laboratory of Genetic Engineering, Institute of Genetics, School of Life Sciences, Fudan University, Shanghai, P.R. China

autor

Li Z.

autor

Wang J.J.

autor

Xu G.I.

autor

Shen Q.

autor

Tian L.

autor

Xue J.I.

autor

Chen J.

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