MBF1 (multi-protein bridging factor 1) is a protein containing a conserved HTH (helix–turn–helix) domain in both eukaryotes and archaea. Eukaryotic MBF1 has been reported to function as a transcriptional co-activator that physically bridges transcription regulators with the core transcription initiation machinery of RNA polymerase II. In addition, MBF1 has been found to be associated with polyadenylated mRNA in yeast as well as in mammalian cells. aMBF1 (archaeal MBF1) is very well conserved among most archaeal lineages; however, its function has so far remained elusive. To address this, we have conducted a molecular characterization of this aMBF1. Affinity purification of interacting proteins indicates that aMBF1 binds to ribosomal subunits. On sucrose density gradients, aMBF1 co-fractionates with free 30S ribosomal subunits as well as with 70S ribosomes engaged in translation. Binding of aMBF1 to ribosomes does not inhibit translation. Using NMR spectroscopy, we show that aMBF1 contains a long intrinsically disordered linker connecting the predicted N-terminal zinc-ribbon domain with the C-terminal HTH domain. The HTH domain, which is conserved in all archaeal and eukaryotic MBF1 homologues, is directly involved in the association of aMBF1 with ribosomes. The disordered linker of the ribosome-bound aMBF1 provides the N-terminal domain with high flexibility in the aMBF1–ribosome complex. Overall, our findings suggest a role for aMBF1 in the archaeal translation process.
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September 2014
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Research Article|
August 07 2014
Archaeal MBF1 binds to 30S and 70S ribosomes via its helix–turn–helix domain
Fabian Blombach;
*Laboratory of Microbiology, Wageningen University, Wageningen 6703HB, The Netherlands
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Helene Launay;
†Institute of Structural and Molecular Biology, University College London (UCL) and Birkbeck College, University of London, London WC1E 6BT, U.K.
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Ambrosius P. L. Snijders;
Ambrosius P. L. Snijders
‡Department of Chemical and Process Engineering, University of Sheffield, Sheffield S1 3JD, U.K.
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Violeta Zorraquino;
Violeta Zorraquino
*Laboratory of Microbiology, Wageningen University, Wageningen 6703HB, The Netherlands
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Hao Wu;
Hao Wu
4
*Laboratory of Microbiology, Wageningen University, Wageningen 6703HB, The Netherlands
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Bart de Koning;
Bart de Koning
*Laboratory of Microbiology, Wageningen University, Wageningen 6703HB, The Netherlands
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Stan J. J. Brouns;
Stan J. J. Brouns
*Laboratory of Microbiology, Wageningen University, Wageningen 6703HB, The Netherlands
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Thijs J. G. Ettema;
Thijs J. G. Ettema
§Department of Cell and Molecular Biology, Science for Life Laboratory, Uppsala University, Uppsala SE-75137, Sweden
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Carlo Camilloni;
Carlo Camilloni
∥Department of Chemistry, University of Cambridge, Cambridge CB2 1EW, U.K.
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Andrea Cavalli;
Andrea Cavalli
∥Department of Chemistry, University of Cambridge, Cambridge CB2 1EW, U.K.
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Michele Vendruscolo;
Michele Vendruscolo
∥Department of Chemistry, University of Cambridge, Cambridge CB2 1EW, U.K.
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Mark J. Dickman;
Mark J. Dickman
‡Department of Chemical and Process Engineering, University of Sheffield, Sheffield S1 3JD, U.K.
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Lisa D. Cabrita;
Lisa D. Cabrita
†Institute of Structural and Molecular Biology, University College London (UCL) and Birkbeck College, University of London, London WC1E 6BT, U.K.
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Anna La Teana;
Anna La Teana
¶Dipartimento di Scienze della Vita e dell’Ambiente, Università Politecnica delle Marche, Ancona 60131, Italy
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Dario Benelli;
Dario Benelli
**Dipartimento Biotecnologie Cellulari ed Ematologia, Università degli Studi di Roma “La Sapienza”, Roma 00161, Italy
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Paola Londei;
Paola Londei
**Dipartimento Biotecnologie Cellulari ed Ematologia, Università degli Studi di Roma “La Sapienza”, Roma 00161, Italy
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John Christodoulou;
John Christodoulou
5
†Institute of Structural and Molecular Biology, University College London (UCL) and Birkbeck College, University of London, London WC1E 6BT, U.K.
5Correspondence may be addressed to either of these authors (email j.christodoulou@ucl.ac.uk or john.vanderoost@wur.nl).
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John van der Oost
John van der Oost
5
*Laboratory of Microbiology, Wageningen University, Wageningen 6703HB, The Netherlands
5Correspondence may be addressed to either of these authors (email j.christodoulou@ucl.ac.uk or john.vanderoost@wur.nl).
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Publisher: Portland Press Ltd
Received:
November 12 2013
Revision Received:
May 13 2014
Accepted:
May 14 2014
Accepted Manuscript online:
May 14 2014
Online ISSN: 1470-8728
Print ISSN: 0264-6021
© The Authors Journal compilation © 2014 Biochemical Society
2014
Biochem J (2014) 462 (2): 373–384.
Article history
Received:
November 12 2013
Revision Received:
May 13 2014
Accepted:
May 14 2014
Accepted Manuscript online:
May 14 2014
Citation
Fabian Blombach, Helene Launay, Ambrosius P. L. Snijders, Violeta Zorraquino, Hao Wu, Bart de Koning, Stan J. J. Brouns, Thijs J. G. Ettema, Carlo Camilloni, Andrea Cavalli, Michele Vendruscolo, Mark J. Dickman, Lisa D. Cabrita, Anna La Teana, Dario Benelli, Paola Londei, John Christodoulou, John van der Oost; Archaeal MBF1 binds to 30S and 70S ribosomes via its helix–turn–helix domain. Biochem J 1 September 2014; 462 (2): 373–384. doi: https://doi.org/10.1042/BJ20131474
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