title: Evaluating Drosophila p53 as a model system for studying cancer mutations.
abstract: The transcription factor p53 is a key tumor suppressor protein. In about half of human cancers, p53 is inactivated directly through mutation in its sequence-specific DNA-binding domain. Drosophila p53 (Dmp53) has similar apoptotic functions as its human homolog and is therefore an attractive model system for studying cancer pathways. To probe the structure and function of Dmp53, we studied the effect of point mutations, corresponding to cancer hot spot mutations in human p53 (Hp53), on the stability and DNA binding affinity of the full-length protein. Despite low sequence conservation, the Hp53 and Dmp53 proteins had a similar melting temperature and generally showed a similar energetic and functional response to cancer-associated mutations. We also found a correlation between the thermodynamic stability of the mutant proteins and their rate of aggregation. The effects of the mutations were rationalized based on homology modeling of the Dmp53 DNA-binding domain, suggesting that the drastically different effects of a cancer mutation in the loop-sheet-helix motif (R282W in Hp53 and R268W in Dmp53) on stability and DNA binding affinity of the two proteins are related to conformational differences in the L1 loop adjacent to the mutation site. On the basis of these data, we discuss the advantages and limitations of using Dmp53 as a model system for studying p53 function and testing p53 rescue drugs.
authors.affiliation: Department of Molecular Microbiology and Biotechnology, George S. Wise Faculty of Life Sciences, Tel Aviv University, Ramat Aviv 69978, Israel.
authors.forename: Gal
authors.initials: G
authors.lastname: Herzog
authors.forename: Andreas C
authors.initials: AC
authors.lastname: Joerger
authors.forename: Merav D
authors.initials: MD
authors.lastname: Shmueli
authors.forename: Alan R
authors.initials: AR
authors.lastname: Fersht
authors.forename: Ehud
authors.initials: E
authors.lastname: Gazit
authors.forename: Daniel
authors.initials: D
authors.lastname: Segal
mesh_terms: D000595:Amino Acid Sequence; D000818:Animals; D004195:Disease Models, Animal; D004330:Drosophila / Q000235:genetics* / Q000378:metabolism; D029721:Drosophila Proteins / Q000737:chemistry* / Q000235:genetics* / Q000378:metabolism; D006801:Humans; D008969:Molecular Sequence Data; D009154:Mutation; D009369:Neoplasms / Q000235:genetics* / Q000378:metabolism; D017434:Protein Structure, Tertiary; D016415:Sequence Alignment; D016159:Tumor Suppressor Protein p53 / Q000737:chemistry* / Q000235:genetics* / Q000378:metabolism
chemical_list: D029721:Drosophila Proteins; C495901:TP53 protein, human; D016159:Tumor Suppressor Protein p53; C474203:p53 protein, Drosophila