Reversible Amyloid Formation by the p53 Tetramerization Domain and a Cancer-associated Mutant

Lee, Amanda S.; Galea, Charles; DiGiammarino, Enrico L.; Jun, Bokkyoo; Murti, Gopal; Ribeiro, Raul C.; Zambetti, Gerard P.; Schultz, Christian; Kriwacki, Richard W.

doi:10.1016/s0022-2836(03)00175-x

Cited by 74 publications

(66 citation statements)

References 40 publications

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“…Heat treatment results in a substantial conformational change consisting of formation of a cross-␤-core and a decrease in ␣-helicity (30). In this respect, the heat-induced transition of Ure2p fibrils is similar to the transitions from a native ␣-helical to ␤-sheet-rich state described previously for other proteins, including rPrP (31,32). On the other hand, our finding emphasizes that substantial conformational rearrangements may occur even after the conversion of ␣-rPrP into mature amyloid structures is completed.…”

Section: Discussionsupporting

confidence: 72%

Annealing Prion Protein Amyloid Fibrils at High Temperature Results in Extension of a Proteinase K-resistant Core

Bocharova

Makarava

Breydo

et al. 2006

Journal of Biological Chemistry

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Amyloids are highly ordered, rigid ␤-sheet-rich structures that appear to have minimal dynamic flexibility in individual polypeptide chains. Here, we demonstrate that substantial conformational rearrangements occur within mature amyloid fibrils produced from full-length mammalian prion protein. The rearrangement results in a substantial extension of a proteinase K-resistant core and is accompanied by an increase in the ␤-sheet-rich conformation. The conformational rearrangement was induced in the presence of low concentrations of Triton X-100 either by brief exposure to 80°C or, with less efficacy, by prolonged incubation at 37°C at pH 7.5 and is referred to here as "annealing." Upon annealing, amyloid fibrils acquired a proteinase K-resistant core identical to that found in bovine spongiform encephalopathy-specific scrapie-associated prion protein. Annealing was also observed when amyloid fibrils were exposed to high temperatures in the absence of detergent but in the presence of brain homogenate. These findings suggest that the amyloid fibrils exist in two conformationally distinct states that are separated by a high energy barrier and that yet unknown cellular cofactors may facilitate transition of the fibrils into thermodynamically more stable state. Our studies provide new insight into the complex behavior of prion polymerization and highlight the annealing process, a previously unknown step in the evolution of amyloid structures.More than 15 severe maladies, including prion, Alzheimer, and Parkinson diseases, are related to the formation of specific ␤-sheet-rich protein aggregates known as amyloid fibrils (1, 2). Recent studies demonstrated that a broad range of proteins unrelated to any known conformational disease are capable of forming ␤-sheet-rich amyloid forms both in vitro and in vivo (3-6). This finding has led to the proposition that the ability to fold into amyloid structures is not a unique property of certain proteins associated with degenerative maladies; rather, it may well be a feature of polypeptides in general (7).Naturally produced amyloid structures are now found in a variety of organisms, including prokaryotes, insects, fish, and mammals (8 -10).Self-assembly of polypeptides into amyloid structures has been implicated in diverse biological functions, including colonization and biofilm formation of Escherichia coli (8), melanosome biogenesis (10), and maintenance of long-term memory (11). Furthermore, amyloid fibrils are utilized as natural biomaterials, e.g. oocyte eggshells in insects and fish (9). The self-propagating conversion of several prion proteins into amyloid forms underlies a non-mendelian type of inheritance in yeast and fungi (12). In additions, the unique physical properties of amyloid structures generated from synthetic peptides have been exploited for creating novel biomaterials and nanodevices (13).Conversion from a native fold into an amyloid state is a complex process that involves several consecutive steps (14). Because small oligomeric aggregates have been shown to b...

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Section: Discussionsupporting

confidence: 72%

Annealing Prion Protein Amyloid Fibrils at High Temperature Results in Extension of a Proteinase K-resistant Core

Bocharova

Makarava

Breydo

et al. 2006

Journal of Biological Chemistry

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“…The TD of p53 has been implicated in multiple aspects of p53 function including sequence-specific DNA binding, subcellular localization, and protein-protein interactions. Disruption of the oligomeric state of p53 can lead to cancer as evidenced by tetramerization-deficient mutations associated with Li-Fraumeni and Li-Fraumenilike syndromes (37) and adrenocortical carcinoma in children (38). Tetramerization domain mediated protein-protein interactions can occur either by direct binding to residues in the TD or indirectly because they necessitate an oligomeric p53 molecule.…”

Section: Discussionmentioning

confidence: 99%

The Conserved CPH Domains of Cul7 and PARC Are Protein-Protein Interaction Modules That Bind the Tetramerization Domain of p53

Kaustov

Lukin

Lemak

et al. 2007

Journal of Biological Chemistry

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Cul7 is a member of the Cullin Ring Ligase (CRL) family and is required for normal mouse development and cellular proliferation. Recently, a region of Cul7 that is highly conserved in the p53-associated, Parkin-like cytoplasmic protein PARC, was shown to bind p53 directly. Here we identify the CPH domains (conserved domain within Cul7, PARC, and HERC2 proteins) of both Cul7 and PARC as p53 interaction domains using size exclusion chromatography and NMR spectroscopy. We present the first structure of the evolutionarily conserved CPH domain and provide novel insight into the Cul7-p53 interaction. The NMR structure of the Cul7-CPH domain reveals a fold similar to peptide interaction modules such as the SH3, Tudor, and KOW domains. The p53 interaction surface of both Cul7 and PARC CPH domains was mapped to a conserved surface distinct from the analogous peptide-binding regions of SH3, KOW, and Tudor domains, suggesting a novel mode of interaction. The CPH domain interaction surface of p53 resides in the tetramerization domain and is formed by residues contributed by at least two subunits.The ubiquitin-proteosome system plays an important role in controlling diverse biological processes, ranging from signal transduction to cell cycle control (1, 2). These complex processes are controlled via specific degradation of individual or groups of proteins. Protein degradation via the ubiquitin pathway involves two successive steps: tagging of the substrate by covalent attachment of multiple ubiquitin molecules (ubiquitylation) and degradation of the tagged protein by 26 S proteosome complex with release of free and reusable ubiquitin (3).Ubiquitylation is the ultimate result of coordinated activity of an enzymatic cascade, which includes a ubiquitin-activating enzyme (E1), 3 a ubiquitin-conjugating enzyme (E2), and ubiquitin-ligating (E3) enzymes. The E3 ligases are the "brain" of this process and determine substrate specificity (4, 5).Cul7, is a recently identified member of the Cullin family of ubiquitin E3 ligases, localizes predominantly in the cytoplasm (6), and forms a unique Skp1-Cul7-Fbx29-like complex with FBXW8, a WD40 containing F-box protein (7-9). Although a target substrate for FBXW8 has not been yet identified, Cul7 recruits RBX1 to form a Skp1-Cul7-Fbx29-like E3 ubiquitin ligase complex (7,8). The biological function of Cul7 is unclear.However, Cul7 appears to play an important role in development (8, 9), and overexpression of Cul7 accelerates the rate of cell proliferation (6).Cul7 has significant sequence similarity (see Fig. 1) with the p53-associated, Parkin-like cytoplasmic protein, PARC (10). Both proteins contain CPH (domain that is conserved in Cul7, PARC, and HERC2 proteins) (11), DOC (DOC1/APC10), and Cullin homology domains (see Fig. 1A) that are linked with E3 ligase function, suggesting that PARC and Cul7 may both function as E3 ubiquitin ligases. PARC has been shown to sequester p53 in the cytoplasm via interaction between the N terminus of PARC and the C terminus of p53 (10). Cul7 may perform f...

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“…This process was found to be enhanced for the " hot-spot" R248Q contact and structural mutant, which is one of many changes in this domain known to destabilize the native structure (47). The other is for the R337H mutation in the tetramerization domain (48,49), which is associated with adrenocortical carcinoma (ACC) in children from southern Brazil (50). Both the WT and mutant peptides formed amyloid-like fibrils when incubated at pH 4.0 and elevated temperatures, with the mutant being susceptible at a lower temperature.…”

Section: Discussionmentioning

confidence: 99%

Unfolding, Aggregation, and Amyloid Formation by the Tetramerization Domain from Mutant p53 Associated with Lung Cancer

et al. 2006

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The p53 tumor suppressor is a tetrameric transcriptional enhancer, and its activity is compromised by mutations that cause amino acid substitutions in its tetramerization domain. Here we analyze the biochemical and biophysical properties of peptides corresponding to amino acids 319 to 358 of wildtype human p53, which includes the tetramerization domain, and that of a cancer-derived mutant with valine substituted for glycine 334. Unlike the wild-type peptide, the G334V peptide forms amyloid fibrils by a two step process under physiological conditions of temperature and pH. Nevertheless, the G334V peptide is capable of forming hetero-oligomers with a wild-type peptide. Computational modeling of the G334V peptide structure suggests that substitution of valine for glycine 334 causes a local distortion that contributes to a β-dominated structural transition leading to amyloid formation. Since the distortion is mostly on the surface, the mutant peptide is still able to form a pseudo-native tetramer complex at higher concentrations and/or lower temperatures. Our † KS was supported in

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Reversible Amyloid Formation by the p53 Tetramerization Domain and a Cancer-associated Mutant

Cited by 74 publications

References 40 publications

Annealing Prion Protein Amyloid Fibrils at High Temperature Results in Extension of a Proteinase K-resistant Core

Annealing Prion Protein Amyloid Fibrils at High Temperature Results in Extension of a Proteinase K-resistant Core

The Conserved CPH Domains of Cul7 and PARC Are Protein-Protein Interaction Modules That Bind the Tetramerization Domain of p53

Unfolding, Aggregation, and Amyloid Formation by the Tetramerization Domain from Mutant p53 Associated with Lung Cancer

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