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Regulation of p53 activity through lysine methylation

Abstract

p53 is a tumour suppressor that regulates the cellular response to genotoxic stresses. p53 is a short-lived protein and its activity is regulated mostly by stabilization via different post-translational modifications. Here we report a novel mechanism of p53 regulation through lysine methylation by Set9 methyltransferase. Set9 specifically methylates p53 at one residue within the carboxyl-terminus regulatory region. Methylated p53 is restricted to the nucleus and the modification positively affects its stability. Set9 regulates the expression of p53 target genes in a manner dependent on the p53-methylation site. The crystal structure of a ternary complex of Set9 with a p53 peptide and the cofactor product S-adenosyl-l-homocysteine (AdoHcy) provides the molecular basis for recognition of p53 by this lysine methyltransferase.

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Figure 1: Set9 methylates p53 in vitro.
Figure 2: Interaction of Set9 with p53 and H3 peptides.
Figure 3: Set9 methylates p53 in vivo.
Figure 4: Set9 potentiates p53 function.
Figure 5: Set9 increases apoptosis in U2OS cells.
Figure 6: Methylation of p53 at the p21 promoter.

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Acknowledgements

We thank L. Vales for comments on the manuscript. We also thank S. L. Berger and members of the Reinberg laboratory, especially A. Kuzmichev and K. Sarma for discussions. We also thank E. White for discussions and P. Chumakov for the gift of the lentivirus LSL–GFP vector, and A. Ivanov for generation of the siRNA-Set9 U2OS cell line. We thank S. Martin for assistance with binding experiments. The GRASP centre at Tufts is acknowledged for providing excellent technical support. This work was supported by a grant from the NIH and the Howard Hughes Medical Institute (D.R.). N.A.B. acknowledges support from the Charlton Award. S.J.G. acknowledges support from the Medical Research Council and from the Association for International Cancer Research.

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Correspondence to Nickolai A. Barlev or Danny Reinberg.

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Supplementary information

Supplementary Figure S1

Set 9 mono-methylates p53. p53 10 mer peptide was methylated by Set9 (52-366aa). (PDF 174 kb)

Supplementary Figure S2

Overlay of cartoon representations of Set9 ternary structures with p53 substrate. (PDF 200 kb)

Supplementary Figure S3

Set 9 regulates expression of BAX and MDM2 genes. (PDF 36 kb)

Supplementary Figure S4

Set9 mediated activation of p21 gene expression requires p53. H1299 cells were stably transfected with Flag-Set9 wt or control vector. (PDF 42 kb)

Supplementary Figure S5

Set9 facilitates apoptosis in p53-dependent manner. (PDF 16 kb)

Supplementary Table S1

Data and refinement statistics for the ternary complex of SET9 with p53(369-374). (PDF 53 kb)

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Chuikov, S., Kurash, J., Wilson, J. et al. Regulation of p53 activity through lysine methylation. Nature 432, 353–360 (2004). https://doi.org/10.1038/nature03117

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