Effect of new olivacine derivatives on p53 protein level

Abstract Background The p53 protein is a transcription factor for many genes, including genes involved in inhibiting cell proliferation and inducing apoptosis in genotoxically damaged and tumor-transformed cells. In more than 55% of cases of human cancers, loss of the essential function of p53 protein is found. In numerous reports, it has been shown that small molecules (chemical compounds) can restore the suppressor function of the mutant p53 protein in tumor cells. The aim of this study was to evaluate the potential anticancer activity of three newly synthesized olivacine derivatives. Methods The study was performed using two cell lines—CCRF/CEM (containing the mutant p53 protein) and A549 (containing a non-mutant, wild-type p53 protein). The cells were incubated with olivacine derivatives for 18 h and then assays were carried out: measurement of the amount of p53 and p21 proteins, detection of apoptosis, cell cycle analysis, and rhodamine 123 accumulation assay (evaluation of P-glycoprotein inhibition). Multiple-criteria decision analysis was used to compare the anticancer activity of the tested compounds. Results Each tested compound caused the reconstitution of suppressor activity of the p53 protein in cells with the mutant protein. In addition, one of the compounds showed significant antitumor activity in both wild-type and mutant cells. For all compounds, a stronger effect on the level of the p53 protein was observed than for the reference compound—ellipticine. Conclusions The observed effects of the tested new olivacine derivatives (pyridocarbazoles) suggest that they are good candidates for new anticancer drugs.

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Functional domains of wild-type and mutant p53 proteins involved in transcriptional regulation, transdominant inhibition, and transformation suppression.

Molecular and Cellular Biology ◽

10.1128/mcb.13.9.5186 ◽

1993 ◽

Vol 13 (9) ◽

pp. 5186-5194 ◽

Cited By ~ 68

Author(s):

T Unger ◽

J A Mietz ◽

M Scheffner ◽

C L Yee ◽

P M Howley

Keyword(s):

Transcriptional Activation ◽

Deletion Mutant ◽

Function Analysis ◽

P53 Protein ◽

Dominant Negative ◽

Terminal Deletion ◽

Mutant P53 ◽

Transactivation Domain ◽

Wild Type ◽

Suppressor Activity

The wild-type (wt) p53 protein has transcriptional activation functions which may be linked to its tumor suppressor activity. Many mutant p53 proteins expressed in cancers have lost the ability to function as transcriptional activators and furthermore may inhibit wt p53 function. To study the mechanisms by which mutant forms of p53 have lost their transactivation function and can act in a dominant negative manner, a structure-function analysis of both mutant and engineered truncated forms of p53 was carried out. We show that different mutant p53 proteins found in cancers vary in the ability to inhibit the transcriptional transactivation and specific DNA binding activities of wt human p53. This transdominant effect was mediated through the carboxy-terminal oligomerization region. The role of the transactivation activity in transformation suppression by wt p53 was also examined by constructing an N-terminal deletion mutant lacking the transactivation domain. This mutant was unable to transactivate but could bind specifically to DNA. Although it was impaired in its ability to suppress transformation of primary rat embryo fibroblasts by adenovirus E1A plus activated ras, the N-terminal deletion mutant still had some suppression activity, suggesting that additional functions of p53 may contribute to transformation suppression.

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Functional domains of wild-type and mutant p53 proteins involved in transcriptional regulation, transdominant inhibition, and transformation suppression

Molecular and Cellular Biology ◽

10.1128/mcb.13.9.5186-5194.1993 ◽

1993 ◽

Vol 13 (9) ◽

pp. 5186-5194

Author(s):

T Unger ◽

J A Mietz ◽

M Scheffner ◽

C L Yee ◽

P M Howley

Keyword(s):

Transcriptional Activation ◽

Deletion Mutant ◽

Function Analysis ◽

P53 Protein ◽

Dominant Negative ◽

Terminal Deletion ◽

Mutant P53 ◽

Transactivation Domain ◽

Wild Type ◽

Suppressor Activity

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Human p53 and CDC2Hs genes combine to inhibit the proliferation of Saccharomyces cerevisiae

Molecular and Cellular Biology ◽

10.1128/mcb.12.3.1357-1365.1992 ◽

1992 ◽

Vol 12 (3) ◽

pp. 1357-1365

Author(s):

J M Nigro ◽

R Sikorski ◽

S I Reed ◽

B Vogelstein

Keyword(s):

Saccharomyces Cerevisiae ◽

Growth Inhibition ◽

Mammalian Cells ◽

P53 Protein ◽

Inducible Promoter ◽

Mutant P53 ◽

Wild Type ◽

Growth Inhibitory Activity ◽

Wild Type P53 ◽

P53 Genes

Human wild-type and mutant p53 genes were expressed under the control of a galactose-inducible promoter in Saccharomyces cerevisiae. The growth rate of the yeast was reduced in cells expressing wild-type p53, whereas cells transformed with mutant p53 genes derived from human tumors were less affected. Coexpression of the normal p53 protein with the human cell cycle-regulated protein kinase CDC2Hs resulted in much more pronounced growth inhibition that for p53 alone. Cells expressing p53 and CDC2Hs were partially arrested in G1, as determined by morphological analysis and flow cytometry. p53 was phosphorylated when expressed in the yeast, but differences in phosphorylation did not explain the growth inhibition attributable to coexpression of p53 and CDC2Hs. These results suggest that wild-type p53 has a growth-inhibitory activity in S. cerevisiae similar to that observed in mammalian cells and suggests that this yeast may provide a useful model for defining the pathways through which p53 acts.

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The mdm-2 oncogene can overcome wild-type p53 suppression of transformed cell growth

Molecular and Cellular Biology ◽

10.1128/mcb.13.1.301-306.1993 ◽

1993 ◽

Vol 13 (1) ◽

pp. 301-306 ◽

Cited By ~ 1

Author(s):

C A Finlay

Keyword(s):

P53 Protein ◽

Mutant P53 ◽

Wild Type ◽

Rat Embryo ◽

Ras Gene ◽

Double Minute ◽

Murine Double Minute ◽

Low Levels ◽

Wild Type P53 ◽

P53 Genes

Expression of a p53-associated protein, Mdm-2 (murine double minute-2), can inhibit p53-mediated transactivation. In this study, overexpression of the Mdm-2 protein was found to result in the immortalization of primary rat embryo fibroblasts (REFs) and, in conjunction with an activated ras gene, in the transformation of REFs. The effect of wild-type p53 on the transforming properties of mdm-2 was determined by transfecting REFs with ras, mdm-2, and normal p53 genes. Transfection with ras plus mdm-2 plus wild-type p53 resulted in a 50% reduction in the number of transformed foci (relative to the level for ras plus mdm-2); however, more than half (9 of 17) of the cell lines derived from these foci expressed low levels of a murine p53 protein with the characteristics of a wild-type p53. These results are in contrast to previous studies which demonstrated that even minimal levels of wild-type p53 are not tolerated in cells transformed by ras plus myc, E1A, or mutant p53. The mdm-2 oncogene can overcome the previously demonstrated growth-suppressive properties of p53.

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TRRAP is essential for regulating the accumulation of mutant and wild-type p53 in lymphoma

Blood ◽

10.1182/blood-2017-09-806679 ◽

2018 ◽

Vol 131 (25) ◽

pp. 2789-2802 ◽

Cited By ~ 9

Author(s):

Alexander Jethwa ◽

Mikołaj Słabicki ◽

Jennifer Hüllein ◽

Marius Jentzsch ◽

Vineet Dalal ◽

...

Keyword(s):

Protein Stability ◽

P53 Protein ◽

Mutant P53 ◽

Wild Type ◽

Therapeutic Targeting ◽

Key Points ◽

Wild Type P53

Key Points The HAT complex member TRRAP is vital for maintaining high p53 levels by shielding it against the natural p53 degradation machinery. Acetylation-modifying complexes regulate p53 protein stability, which may provide a basis for therapeutic targeting of mutant p53.

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CP-31398 Restores DNA-binding Activity to Mutant p53in Vitrobut Does Not Affect p53 Homologs p63 and p73

Journal of Biological Chemistry ◽

10.1074/jbc.m401854200 ◽

2004 ◽

Vol 279 (44) ◽

pp. 45887-45896 ◽

Cited By ~ 45

Author(s):

Mark J. Demma ◽

Serena Wong ◽

Eugene Maxwell ◽

Bimalendu Dasmahapatra

Keyword(s):

Dna Binding ◽

Mammalian Cells ◽

P53 Protein ◽

Binding Activity ◽

Mutant P53 ◽

Dependent Manner ◽

Wild Type ◽

Dna Binding Activity

The p53 protein plays a major role in the maintenance of genome stability in mammalian cells. Mutations of p53 occur in over 50% of all cancers and are indicative of highly aggressive cancers that are hard to treat. Recently, there has been a high degree of interest in therapeutic approaches to restore growth suppression functions to mutant p53. Several compounds have been reported to restore wild type function to mutant p53. One such compound, CP-31398, has been shown effectivein vivo, but questions have arisen to whether it actually affects p53. Here we show that mutant p53, isolated from cells treated with CP-31398, is capable of binding to p53 response elementsin vitro. We also show the compound restores DNA-binding activity to mutant p53 in cells as determined by a chromatin immunoprecipitation assay. In addition, using purified p53 core domain from two different hotspot mutants (R273H and R249S), we show that CP-31398 can restore DNA-binding activity in a dose-dependent manner. Using a quantitative DNA binding assay, we also show that CP-31398 increases significantly the amount of mutant p53 that binds to cognate DNA (Bmax) and its affinity (Kd) for DNA. The compound, however, does not affect the affinity (Kdvalue) of wild type p53 for DNA and only increasesBmaxslightly. In a similar assay PRIMA1 does not have any effect on p53 core DNA-binding activity. We also show that CP-31398 had no effect on the DNA-binding activity of p53 homologs p63 and p73.

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Single chain antibody against the common epitope of mutant p53 restores wild-type activity to mutant p53 protein

FEBS Letters ◽

10.1016/j.febslet.2005.09.031 ◽

2005 ◽

Vol 579 (25) ◽

pp. 5609-5615 ◽

Cited By ~ 2

Author(s):

Sara Orgad ◽

Naomi Goldfinger ◽

Gerald Cohen ◽

Varda Rotter ◽

Beka Solomon

Keyword(s):

P53 Protein ◽

Mutant P53 ◽

Wild Type ◽

Single Chain ◽

Single Chain Antibody ◽

Type Activity ◽

The Common

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Characterization of wild‐type and mutant p53 protein by Raman spectroscopy and multivariate methods

Journal of Raman Spectroscopy ◽

10.1002/jrs.5655 ◽

2019 ◽

Vol 50 (10) ◽

pp. 1388-1394 ◽

Cited By ~ 6

Author(s):

Karen Hernández‐Vidales ◽

Edgar Guevara ◽

Vanesa Olivares‐Illana ◽

Francisco Javier González

Keyword(s):

Raman Spectroscopy ◽

P53 Protein ◽

Mutant P53 ◽

Multivariate Methods ◽

Wild Type

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Wild-type p53 protein potentiates phototoxicity of 2-BA-2-DMHA in HT29 cells expressing endogenous mutant p53

Cancer Letters ◽

10.1016/s0304-3835(99)00013-0 ◽

1999 ◽

Vol 138 (1-2) ◽

pp. 189-195 ◽

Cited By ~ 17

Author(s):

Wen-Geng Zhang ◽

Xiao-Wu Li ◽

Li-Ping Ma ◽

Shen-Wu Wang ◽

Hong-Ying Yang ◽

...

Keyword(s):

P53 Protein ◽

Mutant P53 ◽

Wild Type ◽

Ht29 Cells ◽

Wild Type P53

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Nuclear accumulation of p53 protein is mediated by several nuclear localization signals and plays a role in tumorigenesis

Molecular and Cellular Biology ◽

10.1128/mcb.10.12.6565-6577.1990 ◽

1990 ◽

Vol 10 (12) ◽

pp. 6565-6577

Author(s):

G Shaulsky ◽

N Goldfinger ◽

A Ben-Ze'ev ◽

V Rotter

Keyword(s):

Nuclear Localization ◽

Cell Nucleus ◽

P53 Protein ◽

Simian Virus 40 ◽

T Antigen ◽

Nuclear Accumulation ◽

Mutant P53 ◽

Wild Type ◽

Nuclear Localization Signals ◽

Wild Type P53

The basic carboxy terminus of p53 plays an important role in directing the protein into the nuclear compartment. The C terminus of the p53 molecule contains a cluster of several nuclear localization signals (NLSs) that mediate the migration of the protein into the cell nucleus. NLSI, the most active domain, is highly conserved in genetically diverged species and shares perfect homology with consensus NLS sequences found in other nuclear proteins. The other two NLSs, II and III, appear to be less effective and less conserved. Although nuclear localization is dictated primarily by the NLSs inherent in the primary amino acid sequence, the actual nuclear homing can be modified by interactions with other proteins expressed in the cell. Comparison between wild-type p53 and naturally occurring mutant p53 showed that both protein categories could migrate into the nucleus of rat primary embryonic fibroblasts by essentially similar mechanisms. Nuclear localization of both proteins was totally dependent on the existence of functional NLS domains. In COS cells, however, we found that NLS-deprived wild-type p53 molecules could migrate into the nucleus by complexing with another nuclear protein, simian virus 40 large-T antigen. Wild-type and mutant p53 proteins differentially complexed with viral or cellular proteins, which may significantly affect the ultimate compartmentalization of p53 in the cell; this finding suggests that the actual subcellular compartmentalization of proteins may differ in various cell type milieux and may largely be affected by the ability of these proteins to complex with other proteins expressed in the cell. Experiments designed to test the physiological significance of p53 subcellular localization indicated that nuclear localization of mutant p53 is essential for this protein to enhance the process of malignant transformation of partially transformed cells, suggesting that p53 functions within the cell nucleus.

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