Effects of Denys−Drash Syndrome Point Mutations on the DNA Binding Activity of the Wilms' Tumor Suppressor Protein WT1†

Biochemistry ◽  
1996 ◽  
Vol 35 (37) ◽  
pp. 12070-12076 ◽  
Author(s):  
Franck Borel ◽  
Kathleen C. Barilla ◽  
Tatyana B. Hamilton ◽  
May Iskandar ◽  
Paul J. Romaniuk
Keyword(s):  
Tumor Suppressor ◽  
Dna Binding ◽  
Wilms Tumor ◽  
Point Mutations ◽  
Binding Activity ◽  
Tumor Suppressor Protein ◽  
Dna Binding Activity ◽  
Drash Syndrome

The complex interactions of p53 with target DNA: we learn as we go

2003 ◽  
Vol 81 (3) ◽  
pp. 141-150 ◽  
Author(s):  
Ella Kim ◽  
Wolfgang Deppert
Keyword(s):  
Tumor Suppressor ◽  
Dna Binding ◽  
Target Genes ◽  
Dna Structure ◽  
Structural Features ◽  
Binding Activity ◽  
Tumor Suppressor P53 ◽  
Dna Binding Activity ◽  
Complex Interactions ◽  
Target Dna

The most import biological function of the tumor suppressor p53 is that of a sequence-specific transactivator. In response to a variety of cellular stress stimuli, p53 induces the transcription of an ever-increasing number of target genes, leading to growth arrest and repair, or to apoptosis. Long considered as a "latent" DNA binder that requires prior activation by C-terminal modification, recent data provide strong evidence that the DNA binding activity of p53 is strongly dependent on structural features within the target DNA and is latent only if the target DNA lacks a certain structural signal code. In this review we discuss evidence for complex interactions of p53 with DNA, which are strongly dependent on the dynamics of DNA structure, especially in the context of chromatin. We provide a model of how this complexity may serve to achieve selectivity of target gene regulation by p53 and how DNA structure in the context of chromatin may serve to modulate p53 functions.Key words: tumor suppressor p53, sequence-specific DNA binding, DNA conformation, chromatin, chromatin remodeling.

The CUP2 gene product, regulator of yeast metallothionein expression, is a copper-activated DNA-binding protein

1989 ◽  
Vol 9 (9) ◽  
pp. 4091-4095 ◽  
Author(s):  
C Buchman ◽  
P Skroch ◽  
J Welch ◽  
S Fogel ◽  
M Karin
Keyword(s):  
Dna Binding ◽  
Binding Protein ◽  
Regulatory Gene ◽  
Point Mutations ◽  
Dna Binding Protein ◽  
Binding Activity ◽  
Dna Binding Domain ◽  
Yeast Saccharomyces Cerevisiae ◽  
Dna Binding Activity ◽  
Yeast Strains

CUP2 is a regulatory gene controlling expression of CUP1, which encodes the Cu-binding yeast metallothionein. CUP2, which is identical to the ACE1 gene, encodes a Cu-regulated DNA-binding protein. The CUP2 protein contains a cysteine-rich DNA-binding domain dependent on Cu+ and Ag+ ions which bind the cysteine residues and direct the refolding of the metal-free apoprotein. CUP2 mutant alleles from Cu-sensitive yeast strains have point mutations affecting the DNA-binding activity. These results establish CUP2 as the primary sensor of intracellular Cu+ in the yeast Saccharomyces cerevisiae, functioning as a Cu+-regulated transcriptional activator.

scholarly journals Functional interaction between DP-1 and p53.

1996 ◽  
Vol 16 (10) ◽  
pp. 5888-5895 ◽  
Author(s):  
T S Sørensen ◽  
R Girling ◽  
C W Lee ◽  
J Gannon ◽  
L R Bandara ◽  
...  
Keyword(s):  
Cell Cycle ◽  
Tumor Suppressor ◽  
Dna Binding ◽  
Cellular Proliferation ◽  
Binding Activity ◽  
Terminal Region ◽  
Alternative Mechanism ◽  
Chromosomal Dna ◽  
Dna Binding Activity ◽  
Cell Extracts

The cellular transcription factor DRTF1/E2F and the tumor suppressor protein p53 play important roles in controlling early cell cycle events. DRTF1/E2F is believed to coordinate and integrate the transcription of cell cycle-regulating genes, for example, those involved in DNA synthesis, with the activity of regulatory proteins, such as the retinoblastoma tumor suppressor gene product (pRb), which modulate its transcriptional activity. In contrast, p53 is thought to monitor the integrity of chromosomal DNA and when appropriate interfere with cell cycle progression, for example, in response to DNA damage. Generic DRTF1/E2F DNA binding activity and transcriptional activation arise when members of two distinct families of proteins, such as DP-1 and E2F-1, interact as DP/E2F heterodimers. In many cell types, DP-1 is a widespread component of DRTF1/E2F DNA binding activity which when expressed at high levels oncogenically transforms embryonic fibroblasts. Here, we document an association between DP-1 and p53 and demonstrate its presence in mammalian cell extracts. In vitro p53 interacts with an immunochemically distinct form of DP-1 and in vivo can regulate transcription driven by the DP-1/E2F-1 heterodimer. At the biochemical level, p53 competes with E2F-1 for DP-1, with a consequent reduction in DNA binding activity. Mutational analysis defines within DP-1 a C-terminal region required for the interaction with p53 and within p53 an N-terminal region distinct from that required to bind to MDM2. Our results establish DRTF1/E2F as a common cellular target in growth control mediated through the activities of pRb and p53 and suggest an alternative mechanism through which p53 may regulate cellular proliferation.

scholarly journals p53 regulates cyclophosphamide teratogenesis by controlling caspases 3, 8, 9 activation and NF-κB DNA binding

Reproduction ◽  
2007 ◽  
Vol 134 (2) ◽  
pp. 379-388 ◽  
Author(s):  
Olga Pekar ◽  
Nataly Molotski ◽  
Shoshana Savion ◽  
Amos Fein ◽  
Vladimir Toder ◽  
...  
Keyword(s):  
Cell Proliferation ◽  
Dna Binding ◽  
Molecular Mechanisms ◽  
Binding Activity ◽  
Tumor Suppressor Protein ◽  
Dna Binding Activity ◽  
Organ Level ◽  
Induced Apoptosis ◽  
The Brain ◽  
Transcription Factor Nuclear Factor

The tumor suppressor protein p53 regulates the sensitivity of embryos to such human teratogens as ionizing radiation, diabetes, and cytostatics. Yet, the molecular mechanisms whereby it fulfills this function remain undefined. We used p53 heterozygous (p53+/−) female mice mated with p53+/− males and then exposed to cyclophosphamide (CP) to test whether caspases 3, 8, and 9 and the transcription factor nuclear factor (NF)-κB may serve as p53 targets. Mice were exposed to CP on day 12 of pregnancy and killed on days 15 and 18 of pregnancy to evaluate CP-induced teratogenic effect. The brain and limbs of embryos harvested 24 h after CP treatment were used to evaluate NF-κB (p65) DNA-binding activity by an ELISA-based method, the activity of the caspases by appropriate colorimetric kits, apoptosis, and cell proliferation by TUNEL, and 5′-bromo-2′-deoxyuridine incorporation respectively. We observed that the activation of caspases 3, 8, and 9 and the suppression of NF-κB DNA binding following CP-induced teratogenic insult took place only in teratologically sensitive organs of p53+/+ but not p53−/− embryos. CP-induced apoptosis and suppression of cell proliferation were also more intensive in the former, and they exhibited a higher incidence of structural anomalies, such as open eyes, digit, limb, and tail anomalies. The analysis of the correlations between the p53 embryonic genotype, the activity of the tested molecules, and the CP-induced dysmorphic events at the cellular and organ level suggests caspases 3, 8, and 9 and NF-κB as components of p53-targeting mechanisms in embryos exposed to the teratogen.

scholarly journals DNA-binding activity of the N-terminal cleavage product of poly(ADP-ribose) polymerase is required for UV mediated apoptosis

2000 ◽  
Vol 113 (6) ◽  
pp. 955-961
Author(s):  
J.W. Kim ◽  
J. Won ◽  
S. Sohn ◽  
C.O. Joe
Keyword(s):  
Dna Binding ◽  
Point Mutations ◽  
Binding Activity ◽  
Cleavage Product ◽  
Atp Depletion ◽  
Uv Treatment ◽  
Dna Binding Activity ◽  
Terminal Fragment ◽  
Dna Binding Sites

The role of the N-terminal cleavage product of poly(ADP-ribose) polymerase (PARP) on UV mediated apoptosis was investigated in cultured HeLa cells. Ultrastructural analysis of cells expressing caspase-resistant PARP (PARP(D214A)) revealed the typical features of necrosis following UV treatment. However, cells co-expressing PARP(D214A) with the N-terminal fragment of PARP containing the DNA-binding domain underwent apoptosis instead of necrosis. In this study, we have demonstrated that the DNA-binding activity of the N-terminal fragment of PARP is important for the execution of apoptosis. Point mutations were introduced in the DNA-binding sites of the N-terminal fragment. Cells co-expressing PARP(D214A) with the mutated N-terminal fragments neither stimulated apoptosis nor prevented necrosis in response to UV irradiation. The present study proposes that the DNA-binding activity of the N-terminal fragment of PARP in UV treated cells prevents cellular ATP depletion, a mechanism by which necrotic cell death is triggered.

scholarly journals The CUP2 gene product, regulator of yeast metallothionein expression, is a copper-activated DNA-binding protein.

1989 ◽  
Vol 9 (9) ◽  
pp. 4091-4095 ◽  
Author(s):  
C Buchman ◽  
P Skroch ◽  
J Welch ◽  
S Fogel ◽  
M Karin
Keyword(s):  
Dna Binding ◽  
Binding Protein ◽  
Regulatory Gene ◽  
Point Mutations ◽  
Dna Binding Protein ◽  
Binding Activity ◽  
Dna Binding Domain ◽  
Yeast Saccharomyces Cerevisiae ◽  
Dna Binding Activity ◽  
Yeast Strains

CUP2 is a regulatory gene controlling expression of CUP1, which encodes the Cu-binding yeast metallothionein. CUP2, which is identical to the ACE1 gene, encodes a Cu-regulated DNA-binding protein. The CUP2 protein contains a cysteine-rich DNA-binding domain dependent on Cu+ and Ag+ ions which bind the cysteine residues and direct the refolding of the metal-free apoprotein. CUP2 mutant alleles from Cu-sensitive yeast strains have point mutations affecting the DNA-binding activity. These results establish CUP2 as the primary sensor of intracellular Cu+ in the yeast Saccharomyces cerevisiae, functioning as a Cu+-regulated transcriptional activator.

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