How Do Cancer-Related Mutations Affect the Oligomerisation State of the p53 Tetramerisation Domain?
, Toni Todorovski 1,2,†, Eduard Puig 1, Mireia DÃaz-Lobo 1, Marta Vilaseca 1, Jesús GarcÃa 1, David Andreu 2 and Ernest Giralt 1,3,*Round 1
Reviewer 1 Report
Comments and Suggestions for AuthorsThe authors clearly presented their research on the p53 factor, which is crucial in the process of carcinogenesis. Therefore, the topic is very important. The figures are clear and show the results clearly. The downside is the literature, which is mainly based on articles from 15 - 20 years ago, here you should look for more recent sources. Similarly, the idea of ​​combining results and discussion is also not a good one, a separate section would better highlight the issue. The work is mainly based on the synthesis and analysis of MS and NMR, which is why it is not very diverse. As emphasized by the author, it would require further research. However, it is an interesting work and, with minor adjustments, suitable for the Journal.
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Reviewer 2 Report
Comments and Suggestions for AuthorsOligomerisation or better tetramerization of p53 is crucial for cell growth, cell cycle arrest and induction of apoptosis. Mutation of several TD positions influences tetramerisation. Thus the influence of cancer related mutations on the oligomeric state of p53 TD has been studied. Some of the reported mutations has been incorporated avoiding the flanking region's effect. (T329I, R337H, R342L, L344R, L344P, D352H) and their effect on the secondary and quaternary structures.
The paper reports a well designed research plan , also well executed and the results clearly reported. Thus my opinion is that this paper merits publication,
Some comments I believe would the AAs consider as suggest to implement the paper in some point.
Fig 1 it not easy to identify the red residue with mutation . I suggest the use of a Bold character.
About the use of CD specroscopy. I believe that following the suggestion in ref 53 the difference spectrum would better show the contribution to the spectral dichroic shape at 217nm of the component due to the beta secondary structure. This would explain why, a part the randon coil one) two peptides have a minimum at 10000 while the other peptides at about 20000. The analysis of the biophysical studies reported in supplementary is well done on pH and temperature. This behaviour should be commented.
About NMR I think that would be interesting to discuss the result of the NMR spectrum in S32 that, shifted in the main text, reports the comparison between 37TD-WT blue and 37TD-L344P red. In fact the difference indicate the existence of a strong hydrophobic region around the shifted residues 330,342 and 334.
In the very clever studies of 13C methyl of methionine by HSQC I believe should be important to remember that this methyl is structurally exactly in face of the penylalanine ring 341.Thus a small readjustment of the plane of the ring would influence markedly the HSQC crosspeak with a relevant shift highfield. It is important that the effect is only on the 1H shifts while the 13C is substantially unaffected.This differences in the magnetic effect is well described by Williamson. The other two Phe 328 338 are far probably do not interfere in this affect.. ( see the structure in PDB ID 1OLG –ref 48).
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