Targeting integration of the Saccharomyces Ty5 retrotransposon.

Troy L. Brady, Clarice L. Schmidt, Daniel F. Voytas

Research output: Chapter in Book/Report/Conference proceedingChapter

17 Scopus citations

Abstract

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.

Original languageEnglish (US)
Title of host publicationChromosomal Mitagenesis
PublisherHumana Press
Pages153-163
Number of pages11
Volume435
ISBN (Print)9781588298997
DOIs
StatePublished - Apr 22 2008

Publication series

NameMethods in molecular biology (Clifton, N.J.)
PublisherHumana Press
ISSN (Print)1064-3745

Keywords

  • Chromatin
  • Integrase
  • Integration
  • Retrotransposon
  • Retrovirus
  • Specificity

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    Brady, T. L., Schmidt, C. L., & Voytas, D. F. (2008). Targeting integration of the Saccharomyces Ty5 retrotransposon. In Chromosomal Mitagenesis (Vol. 435, pp. 153-163). (Methods in molecular biology (Clifton, N.J.)). Humana Press. https://doi.org/10.1007/978-1-59745-232-8_11