title: Binding of the termination factor Nsi1 to its cognate DNA site is sufficient to terminate RNA polymerase I transcription in vitro and to induce termination in vivo.
abstract: Different models have been proposed explaining how eukaryotic gene transcription is terminated. Recently, Nsi1, a factor involved in silencing of ribosomal DNA (rDNA), was shown to be required for efficient termination of rDNA transcription by RNA polymerase I (Pol I) in the yeast Saccharomyces cerevisiae. Nsi1 contains Myb-like DNA binding domains and associates in vivo near the 3' end of rRNA genes to rDNA, but information about which and how DNA sequences might influence Nsi1-dependent termination is lacking. Here, we show that binding of Nsi1 to a stretch of 11 nucleotides in the correct orientation was sufficient to pause elongating Pol I shortly upstream of the Nsi1 binding site and to release the transcripts in vitro. The same minimal DNA element triggered Nsi1-dependent termination of pre-rRNA synthesis using an in vivo reporter assay. Termination efficiency in the in vivo system could be enhanced by inclusion of specific DNA sequences downstream of the Nsi1 binding site. These data and the finding that Nsi1 blocks efficiently only Pol I-dependent RNA synthesis in an in vitro transcription system improve our understanding of a unique mechanism of transcription termination.
authors.affiliation: Universität Regensburg, Biochemie Zentrum Regensburg, Lehrstuhl Biochemie III, Regensburg, Germany.
authors.forename: Philipp
authors.initials: P
authors.lastname: Merkl
authors.affiliation: Universität Regensburg, Biochemie Zentrum Regensburg, Lehrstuhl Biochemie III, Regensburg, Germany.
authors.forename: Jorge
authors.initials: J
authors.lastname: Perez-Fernandez
authors.affiliation: Universität Regensburg, Biochemie Zentrum Regensburg, Lehrstuhl Biochemie III, Regensburg, Germany.
authors.forename: Michael
authors.initials: M
authors.lastname: Pilsl
authors.affiliation: Universität Regensburg, Biochemie Zentrum Regensburg, Lehrstuhl Biochemie III, Regensburg, Germany.
authors.forename: Alarich
authors.initials: A
authors.lastname: Reiter
authors.affiliation: Universität Regensburg, Biochemie Zentrum Regensburg, Lehrstuhl Biochemie III, Regensburg, Germany.
authors.forename: Lydia
authors.initials: L
authors.lastname: Williams
authors.affiliation: Universität Regensburg, Biochemie Zentrum Regensburg, Lehrstuhl Biochemie III, Regensburg, Germany.
authors.forename: Jochen
authors.initials: J
authors.lastname: Gerber
authors.affiliation: Universität Regensburg, Biochemie Zentrum Regensburg, Lehrstuhl Biochemie III, Regensburg, Germany.
authors.forename: Maria
authors.initials: M
authors.lastname: Böhm
authors.affiliation: Universität Regensburg, Biochemie Zentrum Regensburg, Lehrstuhl Biochemie III, Regensburg, Germany.
authors.forename: Rainer
authors.initials: R
authors.lastname: Deutzmann
authors.affiliation: Universität Regensburg, Biochemie Zentrum Regensburg, Lehrstuhl Biochemie III, Regensburg, Germany.
authors.forename: Joachim
authors.initials: J
authors.lastname: Griesenbeck
authors.affiliation: Universität Regensburg, Biochemie Zentrum Regensburg, Lehrstuhl Biochemie III, Regensburg, Germany philipp.milkereit@vkl.uni-regensburg.de herbert.tschochner@vkl.uni-regensburg.de.
authors.forename: Philipp
authors.initials: P
authors.lastname: Milkereit
authors.affiliation: Universität Regensburg, Biochemie Zentrum Regensburg, Lehrstuhl Biochemie III, Regensburg, Germany philipp.milkereit@vkl.uni-regensburg.de herbert.tschochner@vkl.uni-regensburg.de.
authors.forename: Herbert
authors.initials: H
authors.lastname: Tschochner
mesh_terms: D001483:Base Sequence; D001665:Binding Sites; D004271:DNA, Fungal / Q000235:genetics / Q000378:metabolism*; D004268:DNA-Binding Proteins / Q000378:metabolism*; D011401:Promoter Regions, Genetic; D011485:Protein Binding; D012318:RNA Polymerase I / Q000378:metabolism*; D012441:Saccharomyces cerevisiae / Q000235:genetics*; D029701:Saccharomyces cerevisiae Proteins / Q000378:metabolism*; D061806:Transcription Termination, Genetic
chemical_list: D004271:DNA, Fungal; D004268:DNA-Binding Proteins; C574300:Nsi1 protein, S cerevisiae; D029701:Saccharomyces cerevisiae Proteins; D012318:RNA Polymerase I
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