Nucleoside-diphosphate kinases (Nme/Nm23/NDPK) constitute a family of evolutionary conserved enzymes involved in many crucial biological processes. The family consists of ten members divided in two groups. Group I, which encompasses Nme1-Nme4, has been extensively studied, especially Nme1 in the context of metastasis formation. The Group II members are evolutionary older, especially the Nme5, Nme6 and Nme7 and little is known about their structure and function. Numerous proteins from evolutionary distinct organisms exhibit extraordinary similarity in primary structure with their orthologues in mammals including humans, as do their predicted secondary and tertiary structures. Therefore, it is presumed that they have similar or identical biochemical and biological functions. Building upon our previous work on the human and sponge Nme family proteins, the proposed project will focus on resolving the structure, as well as biochemical and biological functions of the human Nme6 and its changes during evolution. We will employ a range of biochemical methods and combine them with modern molecular biology methods supported by advanced confocal microscopy techniques.
The main focus of this project is to reveal interactions of p53 with protein partners in melanoma that are capable of modifying its function. We are particularly interested in possible interactions of p53 with family members, namely p53 and p73 isoforms, with nm23, especially nm23-H1 and nm23-H2, and Gli family of proteins.