Reconstitution of the yeast RNA polymerase III transcription system with all recombinant factors

The Journal of Biological Chemistry
Cécile DucrotJoël Acker

Abstract

Transcription factor TFIIIC is a multisubunit complex required for promoter recognition and transcriptional activation of class III genes. We describe here the reconstitution of complete recombinant yeast TFIIIC and the molecular characterization of its two DNA-binding domains, tauA and tauB, using the baculovirus expression system. The B block-binding module, rtauB, was reconstituted with rtau138, rtau91, and rtau60 subunits. rtau131, rtau95, and rtau55 formed also a stable complex, rtauA, that displayed nonspecific DNA binding activity. Recombinant rTFIIIC was functionally equivalent to purified yeast TFIIIC, suggesting that the six recombinant subunits are necessary and sufficient to reconstitute a transcriptionally active TFIIIC complex. The formation and the properties of rTFIIIC-DNA complexes were affected by dephosphorylation treatments. The combination of complete recombinant rTFIIIC and rTFIIIB directed a low level of basal transcription, much weaker than with the crude B'' fraction, suggesting the existence of auxiliary factors that could modulate the yeast RNA polymerase III transcription system.

References

Dec 20, 1992·Journal of Molecular Biology·B R BraunE P Geiduschek
Aug 15, 1991·Proceedings of the National Academy of Sciences of the United States of America·G A KassavetisE P Geiduschek
Oct 10, 1995·Proceedings of the National Academy of Sciences of the United States of America·G A KassavetisM Pisano
Jun 23, 1995·The Journal of Biological Chemistry·N ChaussivertA Sentenac
Dec 6, 1994·Proceedings of the National Academy of Sciences of the United States of America·M C MarsolierA Sentenac
Aug 25, 1994·Nucleic Acids Research·J HuetA Sentenac
Sep 15, 1994·Biochemical and Biophysical Research Communications·S OttonelloG Dieci
May 1, 1993·Molecular and Cellular Biology·J B EschenlauerD A Brow
Nov 14, 1997·Genes & Development·A Ghavidel, M C Schultz
Jun 29, 1999·Cold Spring Harbor Symposia on Quantitative Biology·S ChédinA Sentenac
Nov 24, 1999·Molecular and Cellular Biology·E DeprezA Sentenac
Feb 24, 2000·Nucleic Acids Research·M R Paule, R J White
Jun 23, 2001·Journal of Molecular Biology·E P Geiduschek, G A Kassavetis
Dec 12, 2001·Molecular and Cellular Biology·Hélène Dumay-OdelotChristian Marck
Oct 17, 2002·Genes & Development·Laura Schramm, Nouria Hernandez
Jan 21, 2003·The Journal of Biological Chemistry·Sabine JourdainOlivier Lefebvre
Apr 26, 2003·Trends in Biochemical Sciences·Giorgio Dieci, André Sentenac
Sep 13, 2003·The EMBO Journal·Olivier HarismendyOlivier Lefebvre
Nov 25, 2003·Proceedings of the National Academy of Sciences of the United States of America·Douglas N RobertsBradley R Cairns
Sep 7, 2004·Proceedings of the National Academy of Sciences of the United States of America·Roberto FerrariGiorgio Dieci
Aug 24, 2005·Protein Expression and Purification·Anastasia MylonaChristoph W Müller
Dec 2, 2005·Nature·Jason PtacekMichael Snyder

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Citations

Aug 27, 2009·Proceedings of the National Academy of Sciences of the United States of America·Arounie TavenetJoël Acker
Jul 17, 2008·Genes & Development·Yad Ghavi-HelmJulie Soutourina
Nov 26, 2010·Biochemistry and Cell Biology = Biochimie Et Biologie Cellulaire·Michèle Amouyal
Apr 2, 2014·International Journal of Evolutionary Biology·Sachiko Matsutani
Oct 16, 2012·Biochimica Et Biophysica Acta·Joël AckerOlivier Lefebvre
Nov 13, 2012·Biochimica Et Biophysica Acta·André Sentenac, Michel Riva
Apr 7, 2016·Transcription·L Maximilian Reuter, Katja Sträßer
Nov 10, 2009·Current Opinion in Structural Biology·Michel WernerJulie Soutourina
Jan 24, 2017·Gene·Alain Victor BarrosMarcelo Ricardo Vicari
Jun 22, 2006·The Journal of Biological Chemistry·Elisa GuffantiGiorgio Dieci
Apr 6, 2007·The Journal of Biological Chemistry·Hélène Dumay-OdelotMartin Teichmann
Jun 11, 2015·Nature Communications·Gary MaleChristoph W Müller
Apr 10, 2013·The Journal of Biological Chemistry·Nicholas M I TaylorChristoph W Müller
Nov 1, 2006·Biochemical Society Transactions·G A Kassavetis, E P Geiduschek
May 2, 2018·Mobile DNA·Stephanie CheungVivien Measday
Jul 30, 2015·Genes & Development·L Maximilian ReuterKatja Sträßer

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