A truncation mutant of the 95-kilodalton subunit of transcription factor IIIC reveals asymmetry in Ty3 integration

Molecular and Cellular Biology
Michael AyeSuzanne Sandmeyer

Abstract

Position-specific integration of the retroviruslike element Ty3 near the transcription initiation sites of tRNA genes requires transcription factors IIIB and IIIC (TFIIIB and TFIIIC). Using a genetic screen, we isolated a mutant with a truncated 95-kDa subunit of TFIIIC (TFIIIC95) that reduced the apparent retrotransposition of Ty3 into a plasmid-borne target site between two divergently transcribed tRNA genes. Although TFIIIC95 is conserved and essential, no defect in growth or transcription of tRNAs was detected in the mutant. Steps of the Ty3 life cycle, such as protein expression, proteolytic processing, viruslike particle formation, and reverse transcription, were not affected by the mutation. However, Ty3 integration into a divergent tDNA target occurred exclusively in one orientation in the mutant strain. Investigation of this orientation bias showed that TFIIIC95 and Ty3 integrase interacted in two-hybrid and glutathione S-transferase pulldown assays and that interaction with the mutant TFIIIC95 protein was attenuated. The orientation bias observed here suggests that even for wild-type Ty3, the protein complexes associated with the long terminal repeats are not equivalent in vivo.

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Citations

Jul 9, 2008·Journal of Molecular Medicine : Official Organ of the Gesellschaft Deutscher Naturforscher Und Ärzte·Katrin VoigtZoltán Ivics
Jun 18, 2002·Current Opinion in Cell Biology·Alan M Weiner
Jan 31, 2012·Genome Research·Xiaojie QiSuzanne Sandmeyer
Apr 20, 2005·Genome Research·Becky IrwinSuzanne Sandmeyer
Aug 12, 2005·Cytogenetic and Genome Research·P Lesage, A L Todeschini
Dec 8, 2004·Genetics·Michael AyeSuzanne Sandmeyer
May 7, 2003·Proceedings of the National Academy of Sciences of the United States of America·Suzanne Sandmeyer
May 6, 2003·Proceedings of the National Academy of Sciences of the United States of America·Yunxia ZhuDaniel F Voytas
Jun 28, 2003·Molecular Microbiology·Michael Aye, Suzanne B Sandmeyer
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Jun 30, 2019·Genome Research·Kurt PattersonSuzanne B Sandmeyer
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May 2, 2018·Mobile DNA·Stephanie CheungVivien Measday
Jun 25, 2015·Microbiology Spectrum·Suzanne SandmeyerVirginia Bilanchone

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