Molecular Basis of the Interactions Between the P73 N Terminus and P300: Effects on Transactivation and Modulation by Phosphorylation

Overview

Journal Proc Natl Acad Sci U S A

Specialty Science

Date 2009 Feb 17

PMID 19218448

Citations 23

Authors

Sarah Burge

Daniel P Teufel

Fiona M Townsley

Stefan M V Freund

Mark Bycroft

Alan R Fersht

Affiliations

Soon will be listed here.

Abstract

The transcription factor p73 belongs to the p53 family of proteins and can transactivate a number of target genes in common with p53. Here, we characterized the interaction of the p73 N terminus with four domains of the transcriptional coactivator p300 and with the negative regulator Mdm2 by using biophysical and cellular measurements. We found that, like p53, the N terminus of p73 contained two distinct transactivation subdomains, comprising residues 10-30 and residues 46-67. The p73 N terminus bound weakly to the Taz1, Kix, and IBiD domains of p300 but with submicromolar affinity for Taz2, in contrast to previous reports. We found weaker binding of the p73 N terminus to the p300 domains in vitro correlated with a significant decrease in transactivation activity in a cell line for the QS and T14A mutants, and tighter binding of the phosphomimetic T14D in vitro correlated with an increase in vivo. Further, we found that phosphorylation of T14 increased the affinity of the p73 N terminus for Taz2 10-fold. The phosphomimetic p73alpha T14D caused increased levels of transactivation.

Citing Articles

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References

Weinberg R, Veprintsev D, Fersht A . Cooperative binding of tetrameric p53 to DNA. J Mol Biol. 2004; 341(5):1145-59. DOI: 10.1016/j.jmb.2004.06.071. View

Kussie P, Gorina S, Marechal V, Elenbaas B, Moreau J, Levine A . Structure of the MDM2 oncoprotein bound to the p53 tumor suppressor transactivation domain. Science. 1996; 274(5289):948-53. DOI: 10.1126/science.274.5289.948. View

De Guzman R, Liu H, Dyson H, Wright P . Solution structure of the TAZ2 (CH3) domain of the transcriptional adaptor protein CBP. J Mol Biol. 2000; 303(2):243-53. DOI: 10.1006/jmbi.2000.4141. View

Irwin M, Kondo K, Marin M, Cheng L, Hahn W, Kaelin Jr W . Chemosensitivity linked to p73 function. Cancer Cell. 2003; 3(4):403-10. DOI: 10.1016/s1535-6108(03)00078-3. View

Melino G, De Laurenzi V, Vousden K . p73: Friend or foe in tumorigenesis. Nat Rev Cancer. 2002; 2(8):605-15. DOI: 10.1038/nrc861. View

Campbell K, Lumb K . Structurally distinct modes of recognition of the KIX domain of CBP by Jun and CREB. Biochemistry. 2002; 41(47):13956-64. DOI: 10.1021/bi026222m. View

Teufel D, Freund S, Bycroft M, Fersht A . Four domains of p300 each bind tightly to a sequence spanning both transactivation subdomains of p53. Proc Natl Acad Sci U S A. 2007; 104(17):7009-14. PMC: 1855428. DOI: 10.1073/pnas.0702010104. View

Dumaz N, Milne D, Meek D . Protein kinase CK1 is a p53-threonine 18 kinase which requires prior phosphorylation of serine 15. FEBS Lett. 1999; 463(3):312-6. DOI: 10.1016/s0014-5793(99)01647-6. View

Gaiddon C, Lokshin M, Gross I, Levasseur D, Taya Y, Loeffler J . Cyclin-dependent kinases phosphorylate p73 at threonine 86 in a cell cycle-dependent manner and negatively regulate p73. J Biol Chem. 2003; 278(30):27421-31. DOI: 10.1074/jbc.M300251200. View

10.

Zeng X, Chen L, Jost C, Maya R, Keller D, Wang X . MDM2 suppresses p73 function without promoting p73 degradation. Mol Cell Biol. 1999; 19(5):3257-66. PMC: 84120. DOI: 10.1128/MCB.19.5.3257. View

11.

Ramadan S, Terrinoni A, Catani M, Sayan A, Knight R, Mueller M . p73 induces apoptosis by different mechanisms. Biochem Biophys Res Commun. 2005; 331(3):713-7. DOI: 10.1016/j.bbrc.2005.03.156. View

12.

Hajduk P, Dinges J, Miknis G, Merlock M, Middleton T, Kempf D . NMR-based discovery of lead inhibitors that block DNA binding of the human papillomavirus E2 protein. J Med Chem. 1997; 40(20):3144-50. DOI: 10.1021/jm9703404. View

13.

Zhu J, Zhou W, Jiang J, Chen X . Identification of a novel p53 functional domain that is necessary for mediating apoptosis. J Biol Chem. 1998; 273(21):13030-6. DOI: 10.1074/jbc.273.21.13030. View

14.

Sakaguchi K, Saito S, Higashimoto Y, Roy S, Anderson C, Appella E . Damage-mediated phosphorylation of human p53 threonine 18 through a cascade mediated by a casein 1-like kinase. Effect on Mdm2 binding. J Biol Chem. 2000; 275(13):9278-83. DOI: 10.1074/jbc.275.13.9278. View

15.

Watson I, Blanch A, Lin D, Ohh M, Irwin M . Mdm2-mediated NEDD8 modification of TAp73 regulates its transactivation function. J Biol Chem. 2006; 281(45):34096-103. DOI: 10.1074/jbc.M603654200. View

16.

Malaguarnera R, Vella V, Pandini G, Sanfilippo M, Pezzino V, Vigneri R . TAp73 alpha increases p53 tumor suppressor activity in thyroid cancer cells via the inhibition of Mdm2-mediated degradation. Mol Cancer Res. 2008; 6(1):64-77. DOI: 10.1158/1541-7786.MCR-07-0005. View

17.

Momand J, Zambetti G, Olson D, George D, Levine A . The mdm-2 oncogene product forms a complex with the p53 protein and inhibits p53-mediated transactivation. Cell. 1992; 69(7):1237-45. DOI: 10.1016/0092-8674(92)90644-r. View

18.

Jones E, Dickman M, Whitmarsh A . Regulation of p73-mediated apoptosis by c-Jun N-terminal kinase. Biochem J. 2007; 405(3):617-23. PMC: 2267308. DOI: 10.1042/BJ20061778. View

19.

Chenna R, Sugawara H, Koike T, Lopez R, Gibson T, Higgins D . Multiple sequence alignment with the Clustal series of programs. Nucleic Acids Res. 2003; 31(13):3497-500. PMC: 168907. DOI: 10.1093/nar/gkg500. View

20.

Harms K, Nozell S, Chen X . The common and distinct target genes of the p53 family transcription factors. Cell Mol Life Sci. 2004; 61(7-8):822-42. PMC: 11138747. DOI: 10.1007/s00018-003-3304-4. View