The acidic transcriptional activation domains of VP16 and p53 bind the cellular replication protein A and stimulate in vitro BPV-1 DNA replication

Li, Rong; Botchan, Michael

doi:10.1016/0092-8674(93)90649-b

Cited by 317 publications

(247 citation statements)

References 78 publications

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Section: Sedimentation Equilibrium Studies Of Scrpa Binding To (Dt) 30supporting

confidence: 73%

“…The data in Figure 7A can be globally fit to a two site binding model (see Materials and Methods) yielding macroscopic binding constants for the first (dT) 18 molecule, K 1,obs = (2.1±0.2)×10 7 M −1 and for the second (dT) 18 molecule, K 2,obs = (1.3±0.5)×10 5 M −1 , with corresponding values of quenching, Q 1 =0.23±0.01 and Q 2 =0.33±0.01 (see Table 3). Similar experiments performed with (dT) 16 also show that two moles of (dT) 16 can bind per mole of scRPA at saturation (data not shown).The maximum stoichiometries for the binding of (dT) L as a function of L are plotted in Figure 7C. Clearly, for the binding of (dT) 16 or (dT) 18 , there is sufficient room remaining within the scRPA binding site to form a stable interaction with at least part of a second oligodeoxynucleotide.…”

supporting

confidence: 70%

“…These specificities presumably reflect the fact that RPA proteins also are involved in myriad interactions with other proteins. These include, DNA polymerase α (9), XPA (10), XPG (11), XPF-ERCCI (12), proteins in the Rad52 epistasis group (13), and the Blooms (14) and Werner helicases (15), as well as many others (16)(17)(18)(19).Although SSB proteins from different organisms can differ in architecture, one common feature is that the sites for ssDNA binding consist primarily of so-called oligosaccharide/ oligonucleotide binding folds (OB-folds) (20). Whereas T4 gene 32 protein contains one OBfold, the E. coli SSB tetramer contains four OB-folds and the hetero-trimeric RPA contains six OB-folds.…”

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confidence: 99%

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Saccharomyces cerevisiae Replication Protein A Binds to Single-Stranded DNA in Multiple Salt-Dependent Modes

2006

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We have examined the single stranded DNA binding properties of the S. cerevisiae Replication Protein A (scRPA) using fluorescence titrations, isothermal titration calorimetry and sedimentation equilibrium in order to determine whether scRPA can bind to ssDNA in multiple binding modes. We measured the occluded site size for scRPA binding poly(dT), as well as the stoichiometry, equilibrium binding constants and binding enthalpy of scRPA-((dT) L ) complexes as a function of oligodeoxynucleotide length, L. Sedimentation equilibrium studies show that scRPA is stable heterotrimer over the range of [NaCl] examined (0.02 M to 1.5 M). However, the occluded site size, n, undergoes a salt-dependent transition between values of n=18−20 nucleotides at low [NaCl] to n=26 −28 nucleotides at high [NaCl], with a transition midpoint near 0.36 M NaCl (25.0°C, pH 8.1). Measurements of the stoichiometry of scRPA-(dT) L complexes also show a [NaCl]-dependent change in stoichiometry consistent with the observed change in occluded site size. Measurements of the ΔH obs for scRPA binding to (dT) L at 1.5 M NaCl, yield a contact site size of 28 nucleotides, similar to the occluded site size determined at this [NaCl]. Altogether, these data support a model in which scRPA can bind to ssDNA in at least two binding modes, a low site size mode (n = 18 ± 1 nucleotides), stabilized at low [NaCl], in which only three of its OB-folds are used, and a higher site size mode (n = 27 ± 1 nucleotides), stabilized at higher [NaCl], which uses four of its OB-folds. No evidence for highly cooperative binding of scRPA to ssDNA was found either under any conditions examined. Thus, scRPA shows some similar behavior to the E. coli SSB homo-tetramer, which also shows binding mode transitions, but some significant differences also exist.Single stranded (ss) DNA binding (SSB) proteins exist in nearly all organisms and play central roles in all aspects of DNA metabolism, including DNA replication, repair, and recombination. However, the structural forms of SSB proteins differ considerably among different organisms. For example, the bacteriophage T4 gene 32 protein, the first such SSB protein identified (1, 2) is a monomer, whereas the E. coli SSB protein, is a homotetramer (3,4), as are most SSB proteins from eubacteria. In contrast, the eukaryotic SSB protein, commonly called Replication Protein A (RPA) is a hetero-trimer (5,6).In all eukaryotes, RPA consists of three highly conserved subunits of approximately 70, 32, and 14 kDa, named according to their respective molecular weights, and all three subunits are required for function in vivo (5,6). However, RPA homologues display distinct species specificity, such that scRPA from yeast cannot substitute for human RPA (hsRPA) Figure showing the results of agarose gel electrophoresis experiments at low (20 mM) and high (1.5 M) NaCl concentrations for scRPA-ssDNA complexes formed at different protein to DNA ratios indicating low or no cooperativity upon formation of these complexes. This material is availa...

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Section: Sedimentation Equilibrium Studies Of Scrpa Binding To (Dt) 30supporting

confidence: 73%

supporting

confidence: 70%

mentioning

confidence: 99%

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Saccharomyces cerevisiae Replication Protein A Binds to Single-Stranded DNA in Multiple Salt-Dependent Modes

2006

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“…RPA2 is capable of complex formation with the tumor-suppressor protein p53 modulating the p53-mediated DNA damage checkpoint response by sequestering p53 and preventing the transcriptional activation of genes involved in DNA repair. 26,27 It has been proposed that RPA2 overexpression might effectively inactivate p53, thus allowing cells with damaged DNA to proceed through the cell cycle. 20 Specific interactions have also been demonstrated between RPA2 and menin, another tumor-suppressor protein, whose gene-MEN1-is disrupted in multiple endocrine neoplasia type 1 and which is postulated to participate in the maintenance of genomic integrity.…”

Section: Discussionmentioning

confidence: 99%

“…9 Links between RPA and transcription have been postulated based on the observed interactions between RPA and Gal4, VP16, p53, Stat3, RBT1 and menin. [10][11][12][13][14][15] This protein is present in cells as a heterotrimeric complex consisting of three subunits which, in order of decreasing size, have been designated as RPA1 (70 kDa), RPA2 (32 kDa) and RPA3 (14 kDa). [16][17][18][19] Recently, RPA complex has been identified as an autoantigen in breast cancer patients thus adding another member to the growing list of cancer autoantigens known to play important role in cell proliferation.…”

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confidence: 99%

Replication protein A is an independent prognostic indicator with potential therapeutic implications in colon cancer

et al. 2007

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Replication protein A (RPA), a component of the origin recognition complex, is required for stabilization of single-stranded DNA at early and later stages of DNA replication being thus critical for eukaryotic DNA replication. Experimental studies in colon cancer cell lines have shown that RPA protein may be the target of cytotoxins designed to inhibit cellular proliferation. This is the first study to investigate the expression of RPA1 and RPA2 subunits of RPA protein and assess their prognostic value in colon cancer patients. We analyzed immunohistochemically the expression of RPA1 and RPA2 proteins in a series of 130 colon cancer resection specimens in relation to conventional clinicopathological parameters and patients' survival. Statistical significant positive associations emerged between: (a) RPA1 and RPA2 protein expressions (P ¼ 0.0001), (b) RPA1 and RPA2 labelling indices (LIs) and advanced stage of the disease (P ¼ 0.001 and 0.003, respectively), (c) RPA1 and RPA2 LIs and the presence of lymph node metastasis (P ¼ 0.002 and 0.004, respectively), (d) RPA1 LI and the number of infiltrated lymph nodes (P ¼ 0.021), (e) RPA2 LI and histological grade of carcinomas (P ¼ 0.05). Moreover, a statistical significant higher RPA1 LI was observed in the metastatic sites compared to the original ones (P ¼ 0.012). RPA1 and RPA2 protein expression associated with adverse patients' outcome in both univariate (log rank test: Po0.00001 and 0.00001, respectively) and multivariate (Cox model: P ¼ 0.092 and 0.0001, respectively) statistical analysis. Statistical significant differences according to the expression of RPA1 and RPA2 proteins were also noticed in the survival of stage II (Po0.00001 and 0.0016, respectively) and stage III (P ¼ 0.0029 and 0.0079, respectively) patients. In conclusion, RPA1 and RPA2 proteins appear to be useful prognostic indicators in colon cancer patients and attractive therapeutic targets for regulation by tumor suppressors or other proteins involved in the control of cell proliferation. Modern Pathology (2007) 20, 159-166.

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p53 tumor-suppressor gene: Clues to molecular carcinogenesis

Wang

Harris

1997

J. Cell. Physiol.

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The tumor-suppressor gene product p53 is clearly a component in several biochemical pathways, including transcription, DNA repair, genomic stability, cell-cycle control and apoptosis, that are central to human carcinogenesis. The p53 is functionally inactivated by mutational, viral, and cellular mechanisms in the majority of human cancers. Analysis of the spectrum of p53 mutations provides clues to the etiology and molecular pathogenesis of cancer. Recent insight into the p53-mediated biochemical pathways of cell-cycle arrest and apoptosis has provided further understanding of the mechanisms related to p53-mediated tumor suppression. This insight in turn may provide the potential molecular targets for the development of rational multimodality cancer therapy, including chemo-, immuno-, and gene-therapeutic strategies. The convergence of previously parallel lines of basic, clinical, and epidemiologic investigation may provide an opportunity to transfer research findings rapidly from the laboratory to the clinic.

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The acidic transcriptional activation domains of VP16 and p53 bind the cellular replication protein A and stimulate in vitro BPV-1 DNA replication

Cited by 317 publications

References 78 publications

Saccharomyces cerevisiae Replication Protein A Binds to Single-Stranded DNA in Multiple Salt-Dependent Modes

Saccharomyces cerevisiae Replication Protein A Binds to Single-Stranded DNA in Multiple Salt-Dependent Modes

Replication protein A is an independent prognostic indicator with potential therapeutic implications in colon cancer

p53 tumor-suppressor gene: Clues to molecular carcinogenesis

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