دورية أكاديمية
Targeting integration of the Saccharomyces Ty5 retrotransposon.
| العنوان: | Targeting integration of the Saccharomyces Ty5 retrotransposon. |
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| المؤلفون: | Brady TL; Department of Genetics, Development and Cell Biology, Iowa State University, Ames, IA, USA., Schmidt CL, Voytas DF |
| المصدر: | Methods in molecular biology (Clifton, N.J.) [Methods Mol Biol] 2008; Vol. 435, pp. 153-63. |
| نوع المنشور: | Journal Article; Research Support, N.I.H., Extramural |
| اللغة: | English |
| بيانات الدورية: | Publisher: Humana Press Country of Publication: United States NLM ID: 9214969 Publication Model: Print Cited Medium: Print ISSN: 1064-3745 (Print) Linking ISSN: 10643745 NLM ISO Abbreviation: Methods Mol Biol Subsets: MEDLINE |
| أسماء مطبوعة: | Publication: Totowa, NJ : Humana Press Original Publication: Clifton, N.J. : Humana Press, |
| مواضيع طبية MeSH: | Retroelements/*genetics , Saccharomyces/*genetics, Binding Sites/genetics ; DNA, Fungal/genetics ; DNA, Fungal/metabolism ; Electroporation ; Escherichia coli/genetics ; Integrases/genetics ; Integrases/metabolism ; Plasmids/genetics ; Saccharomyces/metabolism ; Silent Information Regulator Proteins, Saccharomyces cerevisiae/genetics ; Silent Information Regulator Proteins, Saccharomyces cerevisiae/metabolism ; Transformation, Genetic |
| مستخلص: | Many retrotransposons and retroviruses display integration site specificity. Increasingly, this specificity is found to result from recognition by the retroelement of specific chromatin states or DNA-bound protein complexes. A well-studied example of such a targeted retroelement is the Saccharomyces Ty5 retrotransposon, which integrates into heterochromatin at the telomeres and silent mating loci. Targeting is mediated by an interaction between Ty5 integrase (IN) and the heterochromatin protein silent information regulator 4 (Sir4). A small motif of IN, called the targeting domain, is responsible for this interaction. Ty5 integration can be directed to DNA sites outside of heterochromatin by tethering Sir4 to ectopic locations using fusion proteins between Sir4 and a DNA-binding domain. Alternatively, the targeting domain of Ty5 can be swapped with peptides that recognize other protein partners, thereby generating Ty5 elements with new target specificities. The mechanism of Ty5 target site choice suggests that integration specificity of other retrotransposons and retroviruses can be altered by engineering integrases to recognize DNA-bound protein partners. Retroelements can also be used to probe chromatin dynamics and the distribution of protein complexes on chromosomes. Here, we describe the basic assay by which Ty5 integration is monitored to sites of tethered Sir4. |
| معلومات مُعتمدة: | R01 GM61657 United States GM NIGMS NIH HHS |
| المشرفين على المادة: | 0 (DNA, Fungal) 0 (Retroelements) 0 (SIR4 protein, S cerevisiae) 0 (Silent Information Regulator Proteins, Saccharomyces cerevisiae) EC 2.7.7.- (Integrases) |
| تواريخ الأحداث: | Date Created: 20080329 Date Completed: 20080417 Latest Revision: 20080328 |
| رمز التحديث: | 20240628 |
| DOI: | 10.1007/978-1-59745-232-8_11 |
| PMID: | 18370074 |
| قاعدة البيانات: | MEDLINE |
| تدمد: | 1064-3745 |
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| DOI: | 10.1007/978-1-59745-232-8_11 |