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Structure-function analysis of the inverted terminal repeats of the Sleeping Beauty transposon
文献类型：期刊论文
| 作者 | Cui, ZB; Geurts, AM; Liu, GY; Kaufman, CD; Hackett, PB |
| 刊名 | JOURNAL OF MOLECULAR BIOLOGY
 |
| 出版日期 | 2002-05-17 |
| 卷号 | 318期号:5页码:1221-1235 |
| 关键词 | human cells gene therapy HeLa cells vertebrates |
| ISSN号 | 0022-2836 |
| 通讯作者 | Hackett, PB, Discovery Genom Inc, 614 McKinley Pl, Minneapolis, MN 55413 USA |
| 中文摘要 | Translocation 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. |
| 英文摘要 | Translocation 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. |
| WOS标题词 | Science & Technology ; Life Sciences & Biomedicine |
| 学科主题 | Biochemistry & Molecular Biology |
| 类目[WOS] | Biochemistry & Molecular Biology |
| 研究领域[WOS] | Biochemistry & Molecular Biology |
| 关键词[WOS] | IN-VITRO ; CAENORHABDITIS-ELEGANS ; MU-TRANSPOSASE ; HIMAR1-MARINER TRANSPOSON ; CHROMOSOMAL TRANSPOSITION ; TC1-LIKE TRANSPOSONS ; MARINER TRANSPOSASE ; GENE DISRUPTION ; TC1 TRANSPOSON ; DANIO-RERIO |
| 收录类别 | SCI |
| 语种 | 英语 |
| WOS记录号 | WOS:000175767900005 |
| 公开日期 | 2010-10-13 |
| 源URL | [http://ir.ihb.ac.cn/handle/152342/9858]  |
| 专题 | 水生生物研究所_中科院水生所知识产出（2009年前）_期刊论文 |
| 作者单位 | 1.Chinese Acad Sci, Inst Hydrobiol, Wuhan 430072, Peoples R China 2.Discovery Genom Inc, Minneapolis, MN 55413 USA 3.Univ Minnesota, Dept Genet Cell Biol & Dev, St Paul, MN 55108 USA 4.Univ Minnesota, Arnold & Mabel Bechman Ctr Transposon Res, St Paul, MN 55108 USA |
| 推荐引用方式 GB/T 7714 | Cui, ZB,Geurts, AM,Liu, GY,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 | Cui, ZB,Geurts, AM,Liu, GY,Kaufman, CD,&Hackett, PB.(2002).Structure-function analysis of the inverted terminal repeats of the Sleeping Beauty transposon.JOURNAL OF MOLECULAR BIOLOGY,318(5),1221-1235. |
| MLA | Cui, ZB,et al."Structure-function analysis of the inverted terminal repeats of the Sleeping Beauty transposon".JOURNAL OF MOLECULAR BIOLOGY 318.5(2002):1221-1235. |
入库方式： OAI收割
来源：水生生物研究所
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