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Structure-function analysis of the inverted terminal repeats of the Sleeping Beauty transposon
Zongbin Cui; Aron M.Geurts; Geyi Liu; Christopher D.Kaufman; Perry B.Hackett; Hackett, PB, Discovery Genom Inc, 614 McKinley Pl, Minneapolis, MN 55413 USA
2002
Source PublicationJOURNAL OF MOLECULAR BIOLOGY
Volume318Issue:5Pages:1221-1235
AbstractTranslocation of Sleeping Beauty (SB) transposon requires specific binding of SB transposase to inverted terminal repeats (ITRs) of about 230 bp at each end of the transposon, which is followed by a cut-and-paste transfer of the transposon into a target DNA sequence. The ITRs contain two imperfect direct repeats (DRs) of about 32 bp. The outer DRs are at the extreme ends of the transposon whereas the inner DRs are located inside the transposon, 165-166 bp from the outer DRs. Here we investigated the roles of the DR elements in transposition. Although there is a core transposase-binding sequence common to all of the DRs, additional adjacent sequences are required for transposition and these sequences vary in the different DRs. As a result, SB transposase binds less tightly to the outer DRs than to the inner DRs. Two DRs are required in each ITR for transposition but they are not interchangeable for efficient transposition. Each DR appears to have a distinctive role in transposition. The spacing and sequence between the DR elements in an ITR affect transposition rates, suggesting a constrained geometry is involved in the interactions of SB transposase molecules in order to achieve precise mobilization. Transposons are flanked by TA dinucleotide base-pairs that are important for excision; elimination of the TA motif on one side of the transposon significantly reduces transposition while loss of TAs on both flanks of the transposon abolishes transposition. These findings have led to the construction of a more advanced transposon that should be useful in gene transfer and insertional mutagenesis in vertebrates. (C) 2002 Elsevier Science Ltd. All rights reserved.
KeywordHuman Cells Gene Therapy Hela Cells Vertebrates
DepartmentChinese Acad Sci, Inst Hydrobiol, Wuhan 430072, Peoples R China; Discovery Genom Inc, Minneapolis, MN 55413 USA; Univ Minnesota, Dept Genet Cell Biol & Dev, St Paul, MN 55108 USA; Univ Minnesota, Arnold & Mabel Bechman Ctr Transposon Res, St Paul, MN 55108 USA
Subject AreaBiochemistry & Molecular Biology
Indexed BySCI
WOS IDWOS:000175767900005
Citation statistics
Cited Times:171[WOS]   [WOS Record]     [Related Records in WOS]
Document Type期刊论文
Identifierhttp://ir.ihb.ac.cn/handle/152342/9858
Collection期刊论文
Corresponding AuthorHackett, PB, Discovery Genom Inc, 614 McKinley Pl, Minneapolis, MN 55413 USA
Recommended Citation
GB/T 7714
Zongbin Cui,Aron M.Geurts,Geyi Liu,et al. Structure-function analysis of the inverted terminal repeats of the Sleeping Beauty transposon[J]. JOURNAL OF MOLECULAR BIOLOGY,2002,318(5):1221-1235.
APA Zongbin Cui,Aron M.Geurts,Geyi Liu,Christopher D.Kaufman,Perry B.Hackett,&Hackett, PB, Discovery Genom Inc, 614 McKinley Pl, Minneapolis, MN 55413 USA.(2002).Structure-function analysis of the inverted terminal repeats of the Sleeping Beauty transposon.JOURNAL OF MOLECULAR BIOLOGY,318(5),1221-1235.
MLA Zongbin Cui,et al."Structure-function analysis of the inverted terminal repeats of the Sleeping Beauty transposon".JOURNAL OF MOLECULAR BIOLOGY 318.5(2002):1221-1235.
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