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Genetics, Vol. 168, 1159-1176, November 2004, Copyright © 2004
doi:10.1534/genetics.104.028126
Host Factors That Affect Ty3 Retrotransposition in Saccharomyces cerevisiae
Michael Aye*,
Becky Irwin*,
Nadejda Beliakova-Bethell*,
Eric Chen*,
Jennifer Garrus
and
Suzanne Sandmeyer*,1
* Department of Biological Chemistry, University of California College of Medicine, Irvine, California 92697
Department of Biochemistry, University of Utah, Salt Lake City, Utah 84132
1 Corresponding author: Department of Biological Chemistry, Med Sci I D240, University of California College of Medicine, Irvine, CA 92697-1700.
E-mail: sbsandme{at}uci.edu
The retrovirus-like element Ty3 of Saccharomyces cerevisiae integrates at the transcription initiation region of RNA polymerase III. To identify host genes that affect transposition, a collection of insertion mutants was screened using a genetic assay in which insertion of Ty3 activates expression of a tRNA suppressor. Fifty-three loci were identified in this screen. Corresponding knockout mutants were tested for the ability to mobilize a galactose-inducible Ty3, marked with the HIS3 gene. Of 42 mutants tested, 22 had phenotypes similar to those displayed in the original assay. The proteins encoded by the defective genes are involved in chromatin dynamics, transcription, RNA processing, protein modification, cell cycle regulation, nuclear import, and unknown functions. These mutants were induced for Ty3 expression and assayed for Gag3p protein, integrase, cDNA, and Ty3 integration upstream of chromosomal tDNAVal(AAC) genes. Most mutants displayed differences from the wild type in one or more intermediates, although these were typically not as severe as the genetic defect. Because a relatively large number of genes affecting retrotransposition can be identified in yeast and because the majority of these genes have mammalian homologs, this approach provides an avenue for the identification of potential antiviral targets.
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