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Prof. Dr. Wolfgang Seufert


Fakultät für Biologie und Vorklinische Medizin
Institute of Biochemistry, Genetics and Microbiology
Genetik - Cell Cycle Control
Prof. Wolfgang Seufert

Research

 We study mitosis and a new link of mRNA translation to cell proliferation control

Activators of the ubiquitin ligase APC/C
Cyclin-dependent kinases (Cdks) are key regulators of the eukaryotic cell division cycle. Through phosphorylation a various target proteins, cyclinB-Cdk1 promotes early, but blocks late events of mitosis. Therefore, cyclinB proteolysis is important for the completion of nuclear and cell division. Building on work on the ubiquitin system (EMBO J 9:543-50; EMBO J 9:4535-41; TiBS 15:195-8; Nature 373:78-81), we identified a substrate recognition subunit (Hct1, a.k.a. Cdh1) of the anaphase-promoting complex (APC/C) as a regulator of cyclin B proteolysis in budding yeast (Cell 90:683-93; EMBO J 20: 5165-75). Phosphorylation by cyclin B-Cdk1 kinase was found to inhibit the APC/C-Cdh1 ubiquitination pathway (Science 282:1721-4). This antagonism is a fundamental aspect of eukaryotic cell cycle control. Recent work indicated that proteolysis by the APC/C-Cdh1 pathway is restricted to the cell nucleus in budding yeast (MBoC 26:843-58). 
Nuclear localization and APC/C binding of Cdh1 were found to be controlled through separate phosphorylation sites (MBoC 27:2198-212.

 

Research 2016 1

 

Yeast Cdh1 is activated in mitosis by Cdc14 phosphatase, a protein retained in the nucleolus for most of the cell cycle. Current efforts aim to better define the underlying mechanisms, among others by mapping domains of the Net1 nucleolar Cdc14 inhibitor and the related Tof2 protein (Curr Biol 18:1001-5).
 
In another ongoing project we use confocal live cell imaging to study how proteins of the nucleolus are distributed during yeast mitosis. While the rDNA binding protein Fob1 segregated equally, RNA Pol I subunits and ribosome biogenesis factors predominated in the mother cell body, while other nucleolar proteins enriched in the daughter cell. We are various mutants affected in chromosome segregation or Pol I function to understand how this asymmetry in nucleolar protein segregation arises.
 

Cdc123 required for translation initiation and cell cycle entry
The second research focus in the lab concerns Cdc123 and the question how this protein controls entry into the cell division cycle. Cdc123 is conserved among eukaryotes and indispensable for cell viability, but its mechanism of action has remained unknown. Through genetic and biochemical analysis, we found that Cdc123 serves an unanticipated, essential role in the onset of protein synthesis by assembling the translation factor eIF2 (JBC 288:21537-46). This GTP-dependent carrier of the initiator-tRNA is well known also for its regulatory role in the stress-induced reprogramming of gene expression. In a recent collaborative effort, the 3D-structure of Cdc123 was solved (Structure 23:1596-608). The work indicated that Cdc123 is an ATP-binding protein of the ATP-grasp enzyme family. Building on the structural information, we are developing various approaches to work out the molecular mechanism of eIF2 assembly by Cdc123 and the putative regulation of the process. This project is supported by the Deutsche Forschungsgemeinschaft through SFB960.

Research 2016 2

Methods
In our work we are using a range of methods including gene cloning and mutagenesis, PCR-mediated epitope tagging, fluorescence microscopy, flow cytometry, Y2H, Western and co-IP analysis, protein affinity purification, EMSA and ThermoFluor analysis.

Team

Team

Prof. Dr. rer. nat.Wolfgang Seufert
Lehrstuhl für Genetik

Gebäude Westliche Naturwissenschaften D3_2.313
Tel.  0941 943-3162 (Büro)
        0941 943-3075 (Sekretariat)
wolfgang.seufert@ur.de

Dr. rer. nat. Wolfgang Mages
Akademischer Direktor

Gebäude Westliche Naturwissenschaften D3_2.303
Tel. 0941 943-3172
wolfgang.mages@ur.de

Sabine Schneider
Sekretariat Lehrstuhl Genetik

Gebäude Westliche Naturwissenschaften D3_2.311
Tel. 0941 943-3075
sabine.schneider@ur.de

wissenschaftliche Mitarbeiter

Dr. rer. nat. Lea Neumann-Arnold
Postdoc

Gebäude Westliche Naturwissenschaften D3_2.317
Tel. 0941 943-3174 Labor-Durchwahl 3177
lea.neumann-arnold@ur.de

Philipp Girke
Doktorand

Gebäude Westliche Naturwissenschaften D3_2.305
Tel. 0941 943-3180 Labor-Durchwahl 3179
philipp.girke@ur.de

Katharina Hannig
Doktorandin

Gebäude Westliche Naturwissenschaften D3_2.305
Tel. 0941 943-3180 Labor-Durchwahl 3179
katharina.hannig@ur.de

Sebastian Höckner
Doktorand

Gebäude Westliche Naturwissenschaften D3_2.317
Tel. 0941 943-3174 Labor-Durchwahl 3177
sebastian.hoeckner@ur.de

Nina Weber
Doktorandin

Gebäude Westliche Naturwissenschaften D3_2.305
Tel. 0941 943-3180 Labor-Durchwahl 3177
nina-maria.weber@ur.de

technische Mitarbeiter

Andrea Brücher
Technische Assistentin

Gebäude Westliche Naturwissenschaften D3_2.305
Tel. 0941 943-3180 Labor-Durchwahl 3177
andrea.bruecher@ur.de

Antje Machetanz-Morokane
Technische Assistentin

Gebäude Westliche Naturwissenschaften D3_2.303
Tel. 0941 943-3172 Labor-Durchwahl 3179
antje.machetanz-morokane@ur.de

Adelheid Weissgerber
Technische Assistentin

Gebäude Westliche Naturwissenschaften D3_2.303
Tel. 0941 943-3172 Labor-Durchwahl 3179
adelheid.weisgerber@ur.de

Master und Bachelorstudenten


Ehemalige Mitarbeiter

Publications


Höckner S, Neumann-Arnold L, Seufert W. (2016) Dual control by Cdk1 phosphorylation of the budding yeast APC/C ubiquitin ligase activator Cdh1. Mol Biol Cell 27(14):2198-2212.
 
Panvert M, Dubiez E, Arnold L, Perez J, Mechulam Y, Seufert W, Schmitt E. (2015) Cdc123, a Cell Cycle Regulator Needed for eIF2 Assembly, Is an ATP-Grasp Protein with Unique Features. Structure 23(9):1596-608.

Arnold L, Höckner S, Seufert W. (2015) Insights into the cellular mechanism of the yeast ubiquitin ligase APC/C-Cdh1 from the analysis of in vivo degrons. Mol Biol Cell 26(5):843-58.

Perzlmaier AF, Richter F, Seufert W. (2013) Translation initiation requires cell division cycle 123 (Cdc123) to facilitate biogenesis of the eukaryotic initiation factor 2 (eIF2). J Biol Chem. 288(30):21537-46.

Meitinger F, Richter H, Heisel S, Hub B, Seufert W, Pereira G. (2013) A safeguard mechanism regulates Rho GTPases to coordinate cytokinesis with the establishment of cell polarity. PLoS Biol. 11(2):e1001495.

Wittner M, Hamperl S, Stöckl U, Seufert W, Tschochner H, Milkereit P, Griesenbeck J. (2011) Establishment and maintenance of alternative chromatin states at a multicopy gene locus. Cell 145(4):543-54.

Geil, C., Schwab, M., Seufert, W. (2008) A nucleolus-localized activator of Cdc14 phosphatase supports rDNA segregation in yeast mitosis. Current Biology 18, 1001-1005
(PubMed)

Dirscherl, G., Schwab, M., Seufert, W., König, B. (2008) Enhancing the separation of phosphorylated proteins in gel electrophoresis with dinuclear bispyridylmethylamine- tyrosine-acrylamide complexes. Inorganica Chimica Acta, (PubMed)

Willems, A.R., Schwab, M., Tyers, M. (2004) A hitchhiker's guide to the cullin ubiquitin ligases: SCF and its kin. Biochim. Biophys. Acta 1695, 133-170 (PubMed)

Schwab, M., Tyers, M. (2001) Cell cycle. Archipelago of destruction. Nature 413, 268-269 (PubMed)

Schwab, M., Neutzner, M., Möcker, D., Seufert, W. (2001) Yeast Hct1 recognizes the mitotic cyclin Clb2 and other substrates of the ubiquitin ligase APC. EMBO J. 20, 5165-5175 (PubMed)

Menssen, R., Neutzner, A., Seufert, W. (2001) Asymmetric spindle pole localization of yeast Cdc15 kinase links mitotic exit and cytokinesis. Current Biology 11, 345-350 (PubMed)

Zachariae, W., Schwab, M., Nasmyth, K., Seufert, W. (1998) Control of cyclin ubiquitination by CDK-regulated binding of Hct1 to the anaphase promoting complex.  Science 282, 1721-1724 (PubMed)

Schwab, M., Schulze Lutum, A., Seufert, W. (1997) Yeast Hct1 is a regulator of Clb2 cyclin proteolysis.  Cell 90, 683-693 (PubMed)

Betting, J., Seufert, W. (1996) A yeast Ubc9 mutant protein with temperature-sensitive in vivo function is subject to conditional proteolysis by a ubiquitin- and proteasome-dependent pathway. J. Biol. Chem. 271, 25790-25796 (PubMed)

Seufert, W., Futcher, B., Jentsch, S. (1995) Role of a ubiquitin-conjugating enzyme in degradation of S- and M-phase cyclins. Nature 373, 78-81 (PubMed)

Johnson, E., Bartel, B., Seufert, W., Varshavsky, A. (1992) Ubiquitin as a degradation signal. EMBO J. 11, 497-505 (PubMed)

Seufert, W., Jentsch, S. (1992) In vivo function of the proteasome in the ubiquitin pathway. EMBO J. 11, 3077-3080 (PubMed)

Seufert, W., McGrath, J.P., Jentsch, S. (1990) UBC1 encodes a novel member of an essential subfamily of yeast ubiquitin-conjugating enzymes involved in protein degradation. EMBO J. 9, 4535-4541 (PubMed)

Seufert, W., Jentsch, S. (1990) Ubiquitin-conjugating enzymes UBC4 and UBC5 mediate selective degradation of short-lived and abnormal proteins. EMBO J. 9, 543-550 (PubMed)

Seufert,W., Jentsch, S. (1990) Nucleotide sequence of two tRNAArg-tRNAAsp tandem genes linked to duplicated UBC genes in Saccharomyces cerevisiae. Nucleic Acids Res. 18, 1638 (PubMed)

Seufert, W. (1990) Nucleotide sequence of the yeast SDH1 gene encoding a serine dehydratase homolog. Nucleic Acids Res. 18, 3653 (PubMed)

Seufert, W., Lurz, R., Messer, W. (1988) A novel replicon occuring naturally in Escherichia coli is a phage-plasmid hybrid. EMBO J. 7, 4005-4010 (PubMed)

Seufert, W., Dobrinski, B., Lurz, R., Messer, W. (1988) Functionality of the DnaA protein binding site in DNA replication is orientation dependent. J. Biol. Chem. 263, 2719-2723 (PubMed)

Seufert, W., Messer, W. (1987) Start sites for bidirectional in vitro replication inside the replication origin, oriC, of Escherichia coli. EMBO J. 62469-2472 (PubMed)

Seufert, W., Messer, W. (1987) DnaA protein binding to the plasmid origin region can substitute for primosome assembly during replication of pBR322 in vitro. Cell 48, 73-78 (PubMed)

Seufert, W., Messer, W. (1986) Initiation of Escherichia coli minichromosome replication at oriC and protein n’ recognition sites.Two modes for initiating DNA synthesis in EMBO J.5, 3401-3406 (PubMed)

 

 

 

Teaching

CV

2004 Full Professor and Chair at the University of Regensburg, Department of Genetics
1996 - 2004 Associate Professor at the University of Stuttgart,
Institute of industrial Genetics
1994 - 1996 Heisenberg-Fellow and Group Leader at the LMU Munich,
Department of Genetics
1993 - 1994 Research Fellow with Bruce Futcher
at the Cold Spring Harbor Laboratory, New York, USA
1988 - 1993 Post-Doc with Stefan Jentsch
at the Friedrich-Miescher-Laboratory of the
Max-Planck Society in Tübingen, Germany
1985 - 1988 PhD and Diploma Studies with Walter Messer
at the Max-Planck-Institute of Molecular Genetics, Berlin
1981 - 1985 Studies of Biochemistry at the Freie Universität Berlin

Degrees

1995 Habilitation in Genetics at the LMU Munich
1993 Habilitation in Biochemistry at the University of Tübingen
1987 Doctor of Natural Sciences (Dr. rer. nat.) at the FU Berlin

Awards

1994 - 1996 Heisenberg-Fellowship
by the Deutsche Forschungsgemeinschaft
1993 - 1994 Research Fellowship
by the Cold Spring Harbor Laboratory Association


  1. Fakultät für Biologie und Vorklinische Medizin
  2. Faculty Research

Seufert-Lab

 

Seufert191x191

Prof. Dr. Wolfgang Seufert
Tel. 0941/943-3162
wolfgang.seufert@ur.de
Raum: D3_2.313