title: Mod5 protein binds to tRNA gene complexes and affects local transcriptional silencing.
abstract: The tRNA gene-mediated (tgm) silencing of RNA polymerase II promoters is dependent on subnuclear clustering of the tRNA genes, but genetic analysis shows that the silencing requires additional mechanisms. We have identified proteins that bind tRNA gene transcription complexes and are required for tgm silencing but not required for gene clustering. One of the proteins, Mod5, is a tRNA modifying enzyme that adds an N6-isopentenyl adenosine modification at position 37 on a small number of tRNAs in the cytoplasm, although a subpopulation of Mod5 is also found in the nucleus. Recent publications have also shown that Mod5 has tumor suppressor characteristics in humans as well as confers drug resistance through prion-like misfolding in yeast. Here, we show that a subpopulation of Mod5 associates with tRNA gene complexes in the nucleolus. This association occurs and is required for tgm silencing regardless of whether the pre-tRNA transcripts are substrates for Mod5 modification. In addition, Mod5 is bound to nuclear pre-tRNA transcripts, although they are not substrates for the A37 modification. Lastly, we show that truncation of the tRNA transcript to remove the normal tRNA structure also alleviates silencing, suggesting that synthesis of intact pre-tRNAs is required for the silencing mechanism. These results are discussed in light of recent results showing that silencing near tRNA genes also requires chromatin modification.
authors.affiliation: Department of Biological Chemistry, University of Michigan, Ann Arbor, MI 48109-0606, USA.
authors.forename: Matthew
authors.initials: M
authors.lastname: Pratt-Hyatt
authors.forename: Dave A
authors.initials: DA
authors.lastname: Pai
authors.forename: Rebecca A
authors.initials: RA
authors.lastname: Haeusler
authors.forename: Glenn G
authors.initials: GG
authors.lastname: Wozniak
authors.forename: Paul D
authors.initials: PD
authors.lastname: Good
authors.forename: Erin L
authors.initials: EL
authors.lastname: Miller
authors.forename: Ian X
authors.initials: IX
authors.lastname: McLeod
authors.forename: John R
authors.initials: JR
authors.lastname: Yates
authors.forename: Anita K
authors.initials: AK
authors.lastname: Hopper
authors.forename: David R
authors.initials: DR
authors.lastname: Engelke
mesh_terms: D019883:Alkyl and Aryl Transferases / Q000235:genetics / Q000378:metabolism*; D017360:Arabidopsis; D000069059:Atorvastatin; D015152:Blotting, Northern; D002466:Cell Nucleolus / Q000378:metabolism; D047369:Chromatin Immunoprecipitation; D003001:Cloning, Molecular; D017931:DNA Primers / Q000235:genetics; D020868:Gene Silencing / Q000502:physiology*; D006538:Heptanoic Acids; D006801:Humans; D047468:Immunoprecipitation; D017403:In Situ Hybridization; D009841:Oligonucleotides / Q000235:genetics; D011401:Promoter Regions, Genetic / Q000235:genetics / Q000502:physiology; D011758:Pyrroles; D012319:RNA Polymerase II / Q000235:genetics* / Q000502:physiology; D012343:RNA, Transfer / Q000378:metabolism*; D012441:Saccharomyces cerevisiae; D029701:Saccharomyces cerevisiae Proteins / Q000235:genetics / Q000378:metabolism*
chemical_list: D017931:DNA Primers; D006538:Heptanoic Acids; D009841:Oligonucleotides; D011758:Pyrroles; D029701:Saccharomyces cerevisiae Proteins; D012343:RNA, Transfer; D000069059:Atorvastatin; D019883:Alkyl and Aryl Transferases; C068392:MOD5 protein, S cerevisiae; D012319:RNA Polymerase II
keywords: Maf1; RNA silencing
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