Sebastiaan Winkler

Contact

• workRoom C92 Centre for Biomolecular Sciences
  University Park
  Nottingham
  NG7 2RD
  UK
• work0115 8468457
• fax0115 8468877
• sebastiaan.winkler@nottingham.ac.uk

Biography

Following a master's degree in chemistry at the University of Leiden (The Netherlands), I gained a Ph.D. for work on the DNA repair/transcription factor TFIIH under supervision of Prof Jan Hoeijmakers and Dick Bootsma (Erasmus University Rotterdam, The Netherlands). After postdoctoral work at Clare Hall Laboratories, The London Research Institute, Cancer Research UK, and the University Medical Centre Utrecht (The Netherlands), I was appointed to a lectureship in Nottingham in 2007.

Research Summary

We are interested in several aspects of gene regulation in mammalian cells, in particular transcription and mRNA turnover. These processes are essential for normal development and tissue homeostasis,… read more

Recent Publications

• MITTAL, SALONI, ASLAM, AKHMED, DOIDGE, RACHEL, MEDICA, RACHEL and WINKLER, G SEBASTIAAN, 2011. The Ccr4a (CNOT6) and Ccr4b (CNOT6L) deadenylase subunits of the human Ccr4-Not complex contribute to the prevention of cell death and senescence. Molecular biology of the cell. 22(6), 748-58
• ASLAM, AKHMED, MITTAL, SALONI, KOCH, FREDERIC, ANDRAU, JEAN-CHRISTOPHE and WINKLER, G SEBASTIAAN, 2009. The Ccr4-not deadenylase subunits CNOT7 and CNOT8 have overlapping roles and modulate cell proliferation. Molecular biology of the cell. 20(17), 3840-50
• HUANG, ANDING, DE JONG, ROB N, WIENK, HANS, WINKLER, G SEBASTIAAN, TIMMERS, H TH MARC and BOELENS, ROLF, 2009. E2-c-Cbl recognition is necessary but not sufficient for ubiquitination activity. Journal Molecular Biology. 385(2), 507-19
• WINKLER, G.S., 2009. The mammalian anti-proliferative BTG/Tob protein family. Journal of cellular physiology. 222(1), 66-72

• View all publications

Current Research

We are interested in several aspects of gene regulation in mammalian cells, in particular transcription and mRNA turnover. These processes are essential for normal development and tissue homeostasis, and deregulation of gene expression is associated with human disease. Our goal is to understand mechanisms of gene regulation, and to explore this knowledge for potential therapeutic intervention strategies.

The focus of our research is to understand the role of deadenylase enzymes, ribonucleases targeting the poly(A) tails of cytoplasmic mRNA. The human genome encodes approximately ten deadenylase enzymes, of which four are associated with the Ccr4-Not complex. We have used siRNA and genome-wide expression profiling to demonstrate that the highly similar DEDD-type deadenylase subunits Caf1a (CNOT7) and Caf1b (CNOT8) have overlapping roles and regulate cell proliferation of breast cancer cells. We have also investigated the roles of the EEP-type deadenylase subunits of the Ccr4-Not complex, the Ccr4a (CNOT6) and Ccr4b (CNOT6L) paralogues. Using several criteria, including phenotypic and genome-wide expression profiling, we showed that Caf1a/Caf1b and Ccr4a/Ccr4b have distinct roles. As a first step towards exploring the therapeutic potential and identifying small-molecule inhibitors of these enzymes, we have developed a fluorescent assay suitable for small molecule inhibitor screens. Such compounds would be useful research tools and have potential therapeutic use as anti-proliferative reagents for the treatment of cancer, or in stimulating osteoblast activity in osteoporosis and bone regeneration. Finally, we are exploring functional relationships between the Ccr4-Not complex and the BTG/Tob family of anti-proliferative proteins, which are frequently inactivated lung and breast tumours.

We apply a wide range of techniques varying from biochemistry and protein purification to study the basic enzymatic properties of enzymes involved in mRNA turnover in vitro, to cell-based techniques, such as fluorescence microscopy, flow cytometry, and genome-wide expression profiling (Affymetrix GeneChip arrays).

This research is supported by grants from the Association for International Cancer Research (AICR) and the Biotechnology and Biological Sciences Research Council (BBSRC).

Selected publications

Mittal S, Aslam, A, Brierley R, Medica R, Winkler GS (2010) The Ccr4a (CNOT6) and Ccr4b (CNOT6L) deadenylase subunits of the human Ccr4-Not complex are required for the prevention of cell death and senescence. Submitted

Winkler GS (2010) The mammalian anti-proliferative BTG/Tob protein family. J Cell Physiol 222: 66-72.

Aslam A, Mittal S Koch F, Andrau JC, and Winkler GS (2009) The Ccr4-Not deadenylase subunits CNOT7 and CNOT8 have overlapping roles and modulate cell proliferation. Mol Biol Cell 20: 3840-3850.

Winkler GS, Mulder KW, Bardwell VJ, Kalkhoven E, and Timmers HTM (2006) Human Ccr4-Not complex is a ligand-dependent repressor of nuclear receptor-mediated transcription EMBO J 25: 3089-3099.

Teaching

Sterile production and aseptic dispensing (B33F06)

Research projects (B34H08/H09)

• MITTAL, SALONI, ASLAM, AKHMED, DOIDGE, RACHEL, MEDICA, RACHEL and WINKLER, G SEBASTIAAN, 2011. The Ccr4a (CNOT6) and Ccr4b (CNOT6L) deadenylase subunits of the human Ccr4-Not complex contribute to the prevention of cell death and senescence. Molecular biology of the cell. 22(6), 748-58
• ASLAM, AKHMED, MITTAL, SALONI, KOCH, FREDERIC, ANDRAU, JEAN-CHRISTOPHE and WINKLER, G SEBASTIAAN, 2009. The Ccr4-not deadenylase subunits CNOT7 and CNOT8 have overlapping roles and modulate cell proliferation. Molecular biology of the cell. 20(17), 3840-50
• HUANG, ANDING, DE JONG, ROB N, WIENK, HANS, WINKLER, G SEBASTIAAN, TIMMERS, H TH MARC and BOELENS, ROLF, 2009. E2-c-Cbl recognition is necessary but not sufficient for ubiquitination activity. Journal Molecular Biology. 385(2), 507-19
• WINKLER, G.S., 2009. The mammalian anti-proliferative BTG/Tob protein family. Journal of cellular physiology. 222(1), 66-72
• MULDER, KLAAS W, INAGAKI, AKIKO, CAMERONI, ELISABETTA, MOUSSON, FLORENCE, WINKLER, G SEBASTIAAN, DE VIRGILIO, CLAUDIO, COLLART, MARTINE A and TIMMERS, H TH MARC, 2007. Modulation of Ubc4p/Ubc5p-Mediated Stress Responses by the RING-Finger-Dependent Ubiquitin-Protein Ligase Not4p in Saccharomyces cerevisiae. Genetics. 176(1), 181-92
• WINKLER, G.S.., MULDER, K.W., BARDWELL, V.J., KALKHOVEN, E. and TIMMERS, H.T.M., 2006. Human Ccr4-Not complex is a ligand-dependent repressor of nuclear receptor-mediated transcription The EMBO journal. 25(13), 3089-3099
• DREIJERINK, KOEN M A, MULDER, KLAAS W, WINKLER, G SEBASTIAAN, HÖPPENER, JO W M, LIPS, CORNELIS J M and TIMMERS, H TH MARC, 2006. Menin links estrogen receptor activation to histone H3K4 trimethylation. Cancer research. 66(9), 4929-35
• MULDER, KLAAS W, WINKLER, G SEBASTIAAN and TIMMERS, H TH MARC, 2005. DNA damage and replication stress induced transcription of RNR genes is dependent on the Ccr4-Not complex. Nucleic acids research. 33(19), 6384-92
• WINKLER, G SEBASTIAAN and TIMMERS, H TH MARC, 2005. Structure-based approaches to create new e2-e3 enzyme pairs. Methods in enzymology. 399, 355-66
• GILBERT, C., KRISTJUHAN, A., WINKLER, G.S. and SVEJSTRUP, J.Q., 2004. Elongator interactions with nascent mRNA revealed by RNA immunoprecipitation Molecular Cell. 14(4), 457-464
• WINKLER, G SEBASTIAAN, ALBERT, THOMAS K, DOMINGUEZ, CYRIL, LEGTENBERG, YVONNE I A, BOELENS, ROLF and TIMMERS, H TH MARC, 2004. An altered-specificity ubiquitin-conjugating enzyme/ubiquitin-protein ligase pair. Journal of molecular biology. 337(1), 157-65
• DOMINGUEZ, CYRIL, BONVIN, ALEXANDRE M J J, WINKLER, G SEBASTIAAN, VAN SCHAIK, FREDERIK M A, TIMMERS, H TH MARC and BOELENS, ROLF, 2004. Structural model of the UbcH5B/CNOT4 complex revealed by combining NMR, mutagenesis, and docking approaches. Structure (London, England : 1993). 12(4), 633-44
• WINKLER, G.S., KRISTJUHAN, A., ERDJUMENT-BROMAGE, H., TEMPST, P. and SVEJSTRUP, J.Q., 2002. Elongator is a histone H3 and H4 acetyltransferase important for normal histone acetylation levels <i>in vivo</i> Proceedings of the National Academy of Sciences of the United States of America. 99(6), 3517-3522
• SEBASTIAAN WINKLER, G, LACOMIS, LYNNE, PHILIP, JOHN, ERDJUMENT-BROMAGE, HEDIYE, SVEJSTRUP, JESPER Q and TEMPST, PAUL, 2002. Isolation and mass spectrometry of transcription factor complexes. Methods (San Diego, Calif.). 26(3), 260-9
• HAWKES, NICOLA A, OTERO, GABRIEL, WINKLER, G SEBASTIAAN, MARSHALL, NICK, DAHMUS, MICHAEL E, KRAPPMANN, DANIEL, SCHEIDEREIT, CLAUS, THOMAS, CLAIRE L, SCHIAVO, GIAMPIETRO, ERDJUMENT-BROMAGE, HEDIYE and TEM, 2002. Purification and characterization of the human elongator complex. The Journal of biological chemistry. 277(4), 3047-52
• WINKLER, G.S., PETRAKIS, T.G., ETHELBERG, S., TOKUNAGA, M., ERDJUMENT-BROMAGE, H., TEMPST, P. and SVEJSTRUP, J.Q., 2001. RNA polymerase II elongator holoenzyme is composed of two discrete subcomplexes Journal of Biological Chemistry. 276(35), 32743-32749
• WINKLER, G S, SUGASAWA, K, EKER, A P, DE LAAT, W L and HOEIJMAKERS, J H, 2001. Novel functional interactions between nucleotide excision DNA repair proteins influencing the enzymatic activities of TFIIH, XPG, and ERCC1-XPF. Biochemistry. 40(1), 160-5
• WINKLER, G S, ARAÚJO, S J, FIEDLER, U, VERMEULEN, W, COIN, F, EGLY, J M, HOEIJMAKERS, J H, WOOD, R D, TIMMERS, H T and WEEDA, G, 2000. TFIIH with inactive XPD helicase functions in transcription initiation but is defective in DNA repair. The Journal of biological chemistry. 275(6), 4258-66
• SEROZ, T, WINKLER, G S, AURIOL, J, VERHAGE, R A, VERMEULEN, W, SMIT, B, BROUWER, J, EKER, A P, WEEDA, G, EGLY, J M and HOEIJMAKERS, J H, 2000. Cloning of a human homolog of the yeast nucleotide excision repair gene MMS19 and interaction with transcription repair factor TFIIH via the XPB and XPD helicases. Nucleic acids research. 28(22), 4506-13
• WINKLER, G S and HOEIJMAKERS, J H, 1998. From a DNA helicase to brittle hair. Nature genetics. 20(2), 106-7
• WINKLER, G S, VERMEULEN, W, COIN, F, EGLY, J M, HOEIJMAKERS, J H and WEEDA, G, 1998. Affinity purification of human DNA repair/transcription factor TFIIH using epitope-tagged xeroderma pigmentosum B protein. The Journal of biological chemistry. 273(2), 1092-8
• FISCHER, D F, VAN DRUNEN, C M, WINKLER, G S, VAN DE PUTTE, P and BACKENDORF, C, 1998. Involvement of a nuclear matrix association region in the regulation of the SPRR2A keratinocyte terminal differentiation marker. Nucleic acids research. 26(23), 5288-94
• WEEDA, G, DE BOER, J, DONKER, I, DE WIT, J, WINKLER, S B, VAN DER HORST, G T, VERMEULEN, W, BOOTSMA, D and HOEIJMAKERS, J H, 1998. Molecular basis of DNA repair mechanisms and syndromes. Recent results in cancer research. Fortschritte der Krebsforschung. Progrès dans les recherches sur le cancer. 154, 147-55
• VERMEULEN, W, DE BOER, J, CITTERIO, E, VAN GOOL, A J, VAN DER HORST, G T, JASPERS, N G, DE LAAT, W L, SIJBERS, A M, VAN DER SPEK, P J, SUGASAWA, K, WEEDA, G, WINKLER, G S, BOOTSMA, D, EGLY, J M and HOEIJ, 1997. Mammalian nucleotide excision repair and syndromes. Biochemical Society transactions. 25(1), 309-15
• WEEDA, G, ROSSIGNOL, M, FRASER, R A, WINKLER, G S, VERMEULEN, W, VAN 'T VEER, L J, MA, L, HOEIJMAKERS, J H and EGLY, J M, 1997. The XPB subunit of repair/transcription factor TFIIH directly interacts with SUG1, a subunit of the 26S proteasome and putative transcription factor. Nucleic acids research. 25(12), 2274-83