Laboratory of Comparative Enzymology
Head: Wojciech RODE
Staff: Joanna Cieśla, Magdalena Dąbrowska, Adam Jarmuła, Anna Dowierciał (PhD student), Małgorzata Siemiątkowska (PhD student), Piotr Wilk (PhD student), Zbigniew Zieliński
Research profile:
Different aspects of thymidylate biosynthesis, such as its enzymology, structure, regulation, inhibition, as well as post- translational modifications, and their influence on the enzyme properties are investigated. Problems connected with thymi- dylate synthase being a target in chemotherapy, including search for new inhibitors/drugs, drug resistance, specificity and mechanism of action, as well as mechanisms of enzyme inhibition and its specificity, are of particular interest. Another line of studies focuses on cell fate decision mechanisms, using two model experimental systems of senescence-like growth arrest, the nurse cell of Trichinella spiralis parasite and metho- trexate-exposed human colon cancer cells, with bioinformatic approach applied to infer active signaling pathways. Certain molecular aspects of host immunological response to Trichi- nella spiralis infection are also studied.
Current research activities:
• developmental pattern of expression of enzymes involved in nematode thymidylate biosynthesis as potential new target for antiparasitic chemotherapy
• effects of post-translational modifications on thymidylate synthase catalytic and non-catalytic (binding repressing translation of its own mRNA) properties.
• mechanism of thymidylate synthase reaction and its inhi- bition, including search for new inhibitors
• mechanism of establishment of methotrexate–induced cancer cell senescence
Methods:
• classic enzymology, incl. kinetic studies
• genomics and genetic engineering
• purified recombinant enzyme protein production
• enzyme protein chemical modification
• molecular modeling (molecular dynamics, quantum mecha- nics)
• protein crystallography
Selected publications:
Jarmuła A., Frączyk T., Cieplak P., Rode W. (2010) Mechanism of influence of phosphorylation on serine 124 on a decrease of catalytic activity of human thymidylate synthase. Bioorganic & Medicinal Chemistry, 18: 3361-3370.
Świniarska M., Leś A., Rode W., Cieśla J., Millán-Pacheco C., Blake I.O., Pastor N. (2010) Segmental motions of rat thymidylate synthase leading to half-the-sites behaviour. Biopolymers, 93: 549-559.
Dąbrowska M., Mosieniak G., Skierski J., Sikora E., Rode W. (2009) Methotrexate-induced senescence in human adenocarci- noma cells is accompanied by induction of p21waf1/cip1 expression and lack of polyploidy. Cancer Letters, 284: 95-101.
Humeniuk R., Menon L., Mishra P., Gorlick R., Sowers R., Rode W., Pizzorno G., Cheng Y-C., Kemeny N., Bertino J.R., Banerjee D. (2009) Decreased levels of uridine monophosphate kinase as a mechanism of fluoropyrimidine resistance. Molecular Cancer Therapeutics, 8: 1037-1044.
Dąbrowska M., Skoneczny M., Zieliński Z., Rode W. (2008) Nurse cell of Trichinella spp. as a model of long-term cell cycle arrest. Cell Cycle, 7: 2167-2178.
