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2011 | 58 | 1 |

Tytuł artykułu

Sp100 interacts with phage ΦC31 integrase to inhibit its recombination activity

Warianty tytułu

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.

Wydawca

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Rocznik

Tom

58

Numer

1

Opis fizyczny

p.67-73,fig.,ref.

Twórcy

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

  • Calos MP (2006) The phiC31 integrase system for gene therapy. Curr Gene Ther 6: 633-645. 
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  • Chen JZ, Ji CN, Xu GL, Pang RY, Yao JH, Zhu HZ, Xue JL, Jia W (2006) DAXX interacts with phage ΦC31 integrase and inhibits recombination. Nucleic Acids Res 34: 6298-6304. 
  • Ehrhardt A, Engler JA, Xu H, Cherry AM, Kay MA (2006) Molecular analysis of chromosomal rearrangements in mammalian cells after phiC31-mediated integration. Hum Gene Ther 17: 1077-1094. 
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  • Held PK, Olivares EC, Aguilar CP, Finegold M, Calos MP, Grompe M (2005) In vivo correction of murine hereditary tyrosinemia type I by ΦC31 integrase-mediated gene delivery. Mol Ther 11: 399-408. 
  • Keravala A, Ormerod BK, Palmer TD, Calos MP (2008) Long-term transgene expression in mouse neural progenitor cells modified with phiC31 integrase. J Neurosci Methods 173: 299-305. 
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  • Maucksch C, Aneja MK, Hennen E, Bohla A, Hoffmann F, Elfinger M, Rosenecker J, Rudolph C (2008) Cell type differences in activity of the Streptomyces bacteriophage phiC31 integrase. Nucleic Acids Res 36: 5462-5471. 
  • McEwan AR, Rowley PA, Smith MC (2009) DNA binding and synapsis by the large C-terminal domain of phiC31 integrase. Nucleic Acids Res 37: 4764-4773. 
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Typ dokumentu

Bibliografia

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Identyfikator YADDA

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