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Licensed Unlicensed Requires Authentication Published by De Gruyter July 8, 2014

Generic tools for conditionally altering protein abundance and phenotypes on demand

  • Frederik Faden

    Frederik Faden was born in Siegburg, Germany. After a one year stay in Ottawa, Canada he studied biology at the University of Bonn, Germany, from 2005 to 2011, where he finished his diploma on the enzyme ALDEHYDE DEHYDROGENASE 3H1 from Arabidopsis thaliana in the group of Prof. Dr. Dorothea Bartels. Since 2011, he has been a PhD student in the Dissmeyer laboratory at the Leibniz Institute of Plant Biochemistry (IPB) at Halle, Germany, and works on the elucidation of the N-end rule pathway of targeted proteolysis in plants.

    , Stefan Mielke

    Stefan Mielke studies biochemistry at the Martin Luther University of Halle-Wittenberg (MLU), Germany. He received his Bachelors degree in 2012 for investigating the COI1-JAZ co-receptor of jasmonate perception in the group of Prof. Dr. Bettina Hause at the Leibniz Institute of Plant Biochemistry (IPB). Currently, he is working on his Masters thesis at the IPB in Nico Dissmeyer’s lab and is interested in conditional expression of proteins in plants.

    , Dieter Lange

    Dieter Lange studies biochemistry at the MLU and finished his Bachelors degree in Dr. Ute Lechner’s group. He worked on the genetic characterization of microbial communities on artificial beads for waste water purification. He is currently doing his master thesis at the IBP in Nico Dissmeyer’s lab on the optimization of conditional protein depletion in Arabidopsis thaliana.

    and Nico Dissmeyer

    Nico Dissmeyer was born in Detmold, Germany, moved to Tübingen to study biochemistry and philosophy. He moved on via Oregon State University, Corvallis, USA and the Freie Universität Berlin to study biochemistry and biophysics to get his PhD from the Universität zu Köln in 2009 for work on CYCLIN-DEPENDENT-KINASE A;1 at the Max Planck Institute for Plant Breeding Research at Cologne. After a postdoc at the Institut de Biologie Moléculaire des Plantes of the CNRS at Strasbourg, France, he came to the IPB in spring 2011. He is heading the junior research group of the ScienceCampus Halle since October 2011 and interested in highly selective ways for protein recognition and degradation, with special emphasis on the N-end rule pathway.

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From the journal Biological Chemistry

Abstract

Conditional gene expression and modulating protein stability under physiological conditions are important tools in biomedical research. They led to a thorough understanding of the roles of many proteins in living organisms. Current protocols allow for manipulating levels of DNA, mRNA, and of functional proteins. Modulating concentrations of proteins of interest, their post-translational processing, and their targeted depletion or accumulation are based on a variety of underlying molecular modes of action. Several available tools allow a direct as well as rapid and reversible variation right on the spot, i.e., on the level of the active form of a gene product. The methods and protocols discussed here include inducible and tissue-specific promoter systems as well as portable degrons derived from instable donor sequences. These are either constitutively active or dormant so that they can be triggered by exogenous or developmental cues. Many of the described techniques here directly influencing the protein stability are established in yeast, cell culture and in vitro systems only, whereas the indirectly working promoter-based tools are also commonly used in higher eukaryotes. Our major goal is to link current concepts of conditionally modulating a protein of interest’s activity and/or abundance and approaches for generating cell and tissue types on demand in living, multicellular organisms with special emphasis on plants.


Corresponding author: Nico Dissmeyer, Independent Junior Research Group on Protein Recognition and Degradation, Leibniz Institute of Plant Biochemistry (IPB), Weinberg 3, D-06120 Halle/Saale, Germany; and ScienceCampus Halle – Plant-based Bioeconomy, D-06120 Halle/Saale, Germany, e-mail:

About the authors

Frederik Faden

Frederik Faden was born in Siegburg, Germany. After a one year stay in Ottawa, Canada he studied biology at the University of Bonn, Germany, from 2005 to 2011, where he finished his diploma on the enzyme ALDEHYDE DEHYDROGENASE 3H1 from Arabidopsis thaliana in the group of Prof. Dr. Dorothea Bartels. Since 2011, he has been a PhD student in the Dissmeyer laboratory at the Leibniz Institute of Plant Biochemistry (IPB) at Halle, Germany, and works on the elucidation of the N-end rule pathway of targeted proteolysis in plants.

Stefan Mielke

Stefan Mielke studies biochemistry at the Martin Luther University of Halle-Wittenberg (MLU), Germany. He received his Bachelors degree in 2012 for investigating the COI1-JAZ co-receptor of jasmonate perception in the group of Prof. Dr. Bettina Hause at the Leibniz Institute of Plant Biochemistry (IPB). Currently, he is working on his Masters thesis at the IPB in Nico Dissmeyer’s lab and is interested in conditional expression of proteins in plants.

Dieter Lange

Dieter Lange studies biochemistry at the MLU and finished his Bachelors degree in Dr. Ute Lechner’s group. He worked on the genetic characterization of microbial communities on artificial beads for waste water purification. He is currently doing his master thesis at the IBP in Nico Dissmeyer’s lab on the optimization of conditional protein depletion in Arabidopsis thaliana.

Nico Dissmeyer

Nico Dissmeyer was born in Detmold, Germany, moved to Tübingen to study biochemistry and philosophy. He moved on via Oregon State University, Corvallis, USA and the Freie Universität Berlin to study biochemistry and biophysics to get his PhD from the Universität zu Köln in 2009 for work on CYCLIN-DEPENDENT-KINASE A;1 at the Max Planck Institute for Plant Breeding Research at Cologne. After a postdoc at the Institut de Biologie Moléculaire des Plantes of the CNRS at Strasbourg, France, he came to the IPB in spring 2011. He is heading the junior research group of the ScienceCampus Halle since October 2011 and interested in highly selective ways for protein recognition and degradation, with special emphasis on the N-end rule pathway.

Acknowledgments

We thank Maria Klecker and Pavel Reichman for comments on the manuscript. The Independent Junior Research Group is funded by a grant of the ScienceCampus Halle – Plant-based Bioeconomy (SCH) to N.D., a graduate fellowship by the Landesgraduiertenförderung Sachsen-Anhalt to F.F. Support comes from the Deutsche Forschungsgemeinschaft (DFG) Graduate Training Center GRK1026 ‘Conformational Transitions in Macromolecular Interactions’ at Halle, Germany and the Leibniz Institute of Plant Biochemistry (IPB) at Halle, Germany.

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Supplemental Material: The online version of this article (DOI 10.1515/hsz-2014-0160) offers supplementary material, available to authorized users.


Received: 2014-3-4
Accepted: 2014-5-15
Published Online: 2014-7-8
Published in Print: 2014-7-1

©2014 by Walter de Gruyter Berlin/Boston

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