Structures of p63 DNA binding domain in complexes with half-site and with spacer-containing full response elements

Chen, Chen; Gorlatova, Natalia; Kelman, Zvi; Herzberg, Osnat

doi:10.1073/pnas.1013657108

Cited by 42 publications

(65 citation statements)

References 29 publications

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“…The first structure of p53 DBD bound to DNA showed a loop-sheet-helix motif contacting the bases and the phosphate backbone of one quarter-site RE (15). More recent structures of p53 and p63 DBDs in complex with DNA have shown a dimer of DBD dimers where each monomer binds to one of the four basic 5-bp inverted repeat recognition sequences, creating dimerization and tetramerization interfaces (16)(17)(18)(19)(20)(21)(22)(23), but no structure of p73 in complex with DNA is known.…”

mentioning

confidence: 99%

Structure of p73 DNA-binding domain tetramer modulates p73 transactivation

Ethayathulla

Tse

Monti

et al. 2012

Proc. Natl. Acad. Sci. U.S.A.

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The transcription factor p73 triggers developmental pathways and overlaps stress-induced p53 transcriptional pathways. How p53-family response elements determine and regulate transcriptional specificity remains an unsolved problem. In this work, we have determined the first crystal structures of p73 DNA-binding domain tetramer bound to response elements with spacers of different length. The structure and function of the adaptable tetramer are determined by the distance between two half-sites. The structures with zero and one base-pair spacers show compact p73 DNA-binding domain tetramers with large tetramerization interfaces; a two base-pair spacer results in DNA unwinding and a smaller tetramerization interface, whereas a four base-pair spacer hinders tetramerization. Functionally, p73 is more sensitive to spacer length than p53, with one base-pair spacer reducing 90% of transactivation activity and longer spacers reducing transactivation to basal levels. Our results establish the quaternary structure of the p73 DNA-binding domain required as a scaffold to promote transactivation.T he p73 transcription factor that belongs to the p53 protein family and participates in pheromonal sensory, chromosome stability, neurogenesis, inflammation, and osteoblastic differentiation pathways (1, 2). In contrast to p53, p73 is mutated in less than 0.5% of human tumors (3); however, it also participates in p53-dependent and independent pathways, showing oncogenic and tumor suppressor functions (4, 5). These dual opposite activities are due to the presence of two promoters which results in the expression of two main isoforms, TAp73 and ΔNp73 (6).How the members of the p53 protein family trigger different cellular responses still remains an open question. Overall, p73 and p63 can bind to the same p53 response elements (REs), but the activated pathways are different (7,8). There is some redundancy in the activation of stress pathways by the three members of the p53 protein family, but, at the same time, over 100 genes regulated by p73 and p63 are not activated by p53 (9, 10). Like p53, p73 also binds to a 20-bp RE, comprising two half-site decamers in direct orientation that follow a 5′-Pur1-Pur2-Pur3-Cyt4-Ade5/Thy5-Ade6/Thy6-Gua7-Pyr8-Pyr9-Pyr10-3′ consensus sequence (10, 11). Half of the known p53 REs do not have any insertion between the two half-sites and spacers larger than 3 bp are rare, particularly among sites that are transcriptionally activated (12-14). In the case of p53 repressed genes, the cis-element code is poorly defined, but based on a limited number of examples, spacer length appears to be more uniformly distributed and targets have no preference for 0-bp spacers (12).Human p73α is a 636 amino acid protein with a tripartite domain organization similar to its close homolog, p63, and to the shorter 393 amino acid long p53 protein. Members of the p53 family have a disordered N-terminal transactivation domain, a central immunoglobulin-like DNA-binding domain (DBD), and a C terminus that starts with a domain that prom...

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confidence: 99%

Structure of p73 DNA-binding domain tetramer modulates p73 transactivation

Ethayathulla

Tse

Monti

et al. 2012

Proc. Natl. Acad. Sci. U.S.A.

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“…The DBD is a 200-amino acid immunoglobulin-like domain with two ␤-sheets packed as a ␤-sandwich (16). Several structures of the members of the p53 transcription factor family in complex with DNA have been solved (17)(18)(19)(20)(21)(22)(23)(24)(25)(26)(27). The DBD binds as a tetramer to response elements (REs), with each monomer recognizing a 5-bp quartersite of the 20-bp full-site RE.…”

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confidence: 99%

Crystal Structures of the DNA-binding Domain Tetramer of the p53 Tumor Suppressor Family Member p73 Bound to Different Full-site Response Elements

Ethayathulla

Nguyen

Viadiu

2013

Journal of Biological Chemistry

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Background: Members of the p53 protein family bind to full-site response elements (REs) to trigger specific cellular pathways. Results: We solved two crystal structures of the p73 DNA-binding domain in complex with full-site REs. Conclusion: Lys-138 in loop L1 distinguishes between consensus REs. Significance: Conformational changes in Lys-138 might explain specificity between cell arrest and apoptosis target genes.

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“…Docking simulations of p28 and p63 identified motifs within loops connecting the parallel sheet structures of S4, S6, S7, S9, and S10 within the p63 DBD 41 predicted to bind p28 ( Figure 1A). This allowed calculation of the molecular surface and electrostatic potential of the p63 DBD (DeepView ver 4.0.3; Swiss Institute of Bioinformatics, Geneva, Switzerland) ( Figure 1B).…”

Section: Docking Simulation Of P28 and P63 Complexmentioning

confidence: 99%

“…4 Although the DBDs of p63 and p53 share 60% amino acid sequence identity and the overall structure of the p63 DBD closely resembles that of the p53 DBD, the p63 DBD and p53 DBD greatly differ in the binding affinity of their response elements, which likely results from the notable differences in the conformation of the L1 loop. 41,43 This may be the reason that, unlike the p53 DBD/p28 complex ( Figure 1D p28 does not bind to the L1 loop of the p63 DBD. Finally, these predictions clearly suggest that p28 does not inhibit the binding of the RING E3 ligase Pirh2, the HECT E3 ligases AIP4/5, or RACK1 to either p63 or p73.…”

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confidence: 99%

“…DNA fragments of full-length and p63 DBD encoding aa 127-323 of p63 were amplified with specific primer sets, full length; 5′-CGG GAT CCC CAT GAA TTT TGA AAC TTC ACG GTG TGC C-3′ and 5′-ATA GTC GAC TCA CTC CCC CTC CTC TTT GAT GC-3′, p63DBD; 5′-CGG GAT CCC CTC CAA CAC CGA CTA CCC AGG CCC GC-3′ and 5′-ATA GTC GAC TCA CTG CTT TCT GAT GCT ATC TTC ATC-3′ containing the BamHI and Sal I restriction enzyme sites, respectively. 41 DNA fragments were introduced into the pGEX5X3 plasmid DNA in frame. Glutathione S-transferase (GST)-fusion proteins were purified by Glutathione Sepharose 4B affinity chromatography and HiLoad™ Superdex 75 gel-filtration column connected to the ÄKTA prime plus FPLC system (GE Healthcare, Little Chalfont, UK).…”

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A nanotechnological, molecular-modeling, and immunological approach to study the interaction of the anti-tumorigenic peptide p28 with the p53 family of proteins

Coppari¹,

Yamada²,

Bizzarri³

et al. 2014

IJN

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p28 is an anionic, amphipathic, cell-penetrating peptide derived from the cupredoxin azurin that binds to the DNA-binding domain (DBD) of the tumor suppressor protein, p53, and induces a post-translational increase in the level of wild type and mutated p53 in a wide variety of human cancer cells. As p63 and p73, additional members of the p53 superfamily of proteins, also appear to be involved in the cellular response to cancer therapy and are reportedly required for p53-induced apoptosis, we asked whether p28 also binds to p63 and p73. Atomic force spectroscopy demonstrates that p28 forms a stable, high-affinity complex with full-length p63, the DBD of p63, and full-length p73. Exposure to p28 decreased the level of TAp63α and ΔNp63α, the truncated form of p63, in p53 wild type and mutated human breast cancer cells, respectively. p28 increased the level of TAp73α, but not ΔNp73α, in the same breast cancer cell lines. In contrast, p28 increased the level of the TA and ΔN isoforms of p63 in p53 wild type, but not in p53 mutated melanoma cells, while decreasing TA p73α in p53 wild type and mutated human melanoma cells. All changes were mirrored by an associated change in the expression of the HECT E3 ligases Itch/AIP4, AIP5, and the RING E3 ligase Pirh2, but not in the receptor for activated C kinase or the RING E3 ligases Mdm2 and Cop1. Collectively, the data suggest that molecules such as p28 bind with high affinity to the DBD of p63 and p73 and alter their expression independent of the Mdm2 and Cop1 pathways.

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Structures of p63 DNA binding domain in complexes with half-site and with spacer-containing full response elements

Cited by 42 publications

References 29 publications

Structure of p73 DNA-binding domain tetramer modulates p73 transactivation

Structure of p73 DNA-binding domain tetramer modulates p73 transactivation

Crystal Structures of the DNA-binding Domain Tetramer of the p53 Tumor Suppressor Family Member p73 Bound to Different Full-site Response Elements

A nanotechnological, molecular-modeling, and immunological approach to study the interaction of the anti-tumorigenic peptide p28 with the p53 family of proteins

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