Role In DNA Repair Pathways Research Articles

BRCA1 mRNA expression and response in locally advanced esophageal cancer patients (p) treated with chemoradiotherapy.

e14573 Background: Chemoradiotherapy is an effective treatment in esophageal cancer p; however, there are no reliable pretreatment clinical parameters to predict the efficacy of this multimodal approach. BRCA1 plays a crucial role in DNA repair pathways, and decreased BRCA1 mRNA expression has been associated with sensitivity to cisplatin and to radiotherapy in several tumors. Methods: We analyzed BRCA1 mRNA expr ession by quantitative PCR in pretreatment endoscopic biopsies from 43 locally advanced esophageal cancer p treated with cisplatin plus fluorouracil and concurrent with radiotherapy, followed by esophagectomy when feasible. p were divided into groups according to tercil es of BRCA1 expression levels. Since the sample size was relatively small, outcomes for those in the lowest tercile were compared to those in the intermediate and high est tercil es together. Results: Eight of 43 p were classified as adenocarcinomas and 35 as squamous cell carcinomas. Nine p had stage II and 34 stage III disease. Of 23 p who underwent esophagectomy after chemoradiotherapy, 10 (43.5%) attained a significant pathological r esponse (pR) (pT0-2N0M0). Of 6 p in the lowest tercile of BRCA1 undergoing surgery, 4 (66%) had a significant pR, while of 17 p in the intermediate and highest tercil es, only 6 (36%) attained a significant pR (p = 0.3). 5-year survival was 18% for all 43 p, 21% for p in the low est tercile of BRCA1, and 17% for p in the intermediate and highest terciles (p = 0.8). Conclusions: Low BRCA1 expr ession is associated with a trend towards better pR in p treated with preoperative chemoradiotherapy. The fact that this association did not reach significance may be due to the small number of patients, and additional patients are currently being included in this analysis to further confirm these results. No significant financial relationships to disclose.

Structural characterization of BRCT–tetrapeptide binding interactions

BRCT(BRCA1) plays a major role in DNA repair pathway, and does so by recognizing the conserved sequence pSXXF in its target proteins. Remarkably, tetrapeptides containing pSXXF motif bind with high specificity and micromolar affinity. Here, we have characterized the binding interactions of pSXXF tetrapeptides using NMR spectroscopy and calorimetry. We show that BRCT is dynamic and becomes structured on binding, that pSer and Phe residues dictate overall binding, and that the binding affinities of the tetrapeptides are intimately linked to structural and dynamic changes both in the BRCT(BRCA1) and tetrapeptides. These results provide critical insights for designing high-affinity BRCT(BRCA1) inhibitors.

Platinum Resistance: The Role of DNA Repair Pathways

Although platinum chemotherapeutic agents such as carboplatin, cisplatin, and oxaliplatin are used to treat a broad range of malignant diseases, their efficacy in most cancers is limited by the development of resistance. There are multiple factors that contribute to platinum resistance but alterations of DNA repair processes have been known for some time to be important in mediating resistance. Recently acquired knowledge has provided insight into the molecular mechanisms of DNA repair pathways and their effect on response to chemotherapy. This review will discuss the most important DNA repair pathways known to be involved in the platinum response, i.e., nucleotide excision repair (NER) and mismatch repair (MMR), and will briefly touch on the role of BRCA in DNA repair. The therapeutic implications of alterations in DNA repair which affect response to platinum in the treatment of patients with malignant disease, such as excision repair cross-complementation group 1 (ERCC1) deficiency and mismatch repair deficiency, will be reviewed.

Repair of Damaged Bases

Repair of Damaged Bases

EFFECTS OF UV‐B‐INDUCED DNA DAMAGE AND PHOTOINHIBITION ON GROWTH OF TEMPERATE MARINE RED MACROPHYTES: HABITAT‐RELATED DIFFERENCES IN UV‐B TOLERANCE

The sensitivity to UV‐B radiation (UVBR: 280–315 nm) was tested for littoral (Palmaria palmata[L.] O. Kuntze, Chondrus crispus Stackhouse) and sublittoral (Phyllophora pseudoceranoides S. G. Gmelin, Rhodymenia pseudopalmata[Lamouroux] Silva, Phycodrys rubens[L.] Batt, Polyneura hilliae[Greville] Kylin) red macrophytes from Brittany, France. Algal fragments were subjected to daily repeated exposures of artificial UVBR that were realistic for springtime solar UVBR at the water surface in Brittany. Growth, DNA damage, photoinhibition, and UV‐absorbing compounds were monitored during 2 weeks of PAR + UV‐A radiation (UVAR) + UVBR, whereas PAR + UVAR and PAR treatments were used as controls. The littoral species showed a higher UV tolerance than the sublittoral species. After 2 weeks, growth of P. palmata and C. crispus was not significantly affected by UVBR, and DNA damage, measured as the number of cyclobutane‐pyrimidine dimers per 106 nucleotides, was negligible. Photoinhibition, determined as the decline in optimal quantum yield, was low and decreased during the course of the experiment, coinciding with the production of UV‐absorbing compounds in these species. In contrast, no UV‐absorbing compounds were induced in the sublittoral species. Growth rates of P. pseudoceranoides and R. pseudopalmata were reduced by 40% compared with the PAR treatment. Additionally, constant levels of DNA damage and pronounced photoinhibition were observed after the UVBR treatments. Growth was completely halted for Phycodrys rubens and Polyneura hilliae, whereas DNA damage accumulated in the course of the experiment. Because Phycodrys rubens and Polyneura hilliae showed the same degree of photoinhibition as the other sublittoral species, it appears that the accumulation of DNA damage may have been responsible for the complete inhibition of growth. The results suggest an important role of DNA repair pathways in determining the UV sensitivity in red macrophytes.

Defective nucleotide excision repair in Xpc mutant mice and its association with cancer predisposition

Mice that are genetically engineered are becoming increasingly more powerful tools for understanding the molecular pathology of many human hereditary diseases, especially those that confer an increased predisposition to cancer. We have generated mouse strains defective in the Xpc gene, which is required for nucleotide excision repair (NER) of DNA. Homozygous mutant mice are highly prone to skin cancer following exposure to UVB radiation, and to liver and lung cancer following exposure to the chemical carcinogen acetylaminofluorene (AAF). Skin cancer predisposition is significantly augmented when mice are additionally defective in Trp53 (p53) gene function. We also present the results of studies with mice that are heterozygous mutant in the Apex (Hap1, Ref-1) gene required for base excision repair and with mice that are defective in the mismatch repair gene Msh2. Double and triple mutant mice mutated in multiple DNA repair genes have revealed several interesting overlapping roles of DNA repair pathways in the prevention of mutation and cancer.

Biochemical and Genetic Defects in the DNA-Dependent Protein Kinase in Murine scid Lymphocytes

The scid gene product has been identified as the 460-kDa catalytic subunit of the DNA-dependent protein kinase (DNA-PKcs p460), a member of the phosphatidylinositol 3-kinase family. DNA-PK activity is undetectable in scid cells, but the molecular basis for this defect has not been identified. Here we report that expression of p460 in scid lymphocyte precursors is detectable but is reduced at least 10-fold relative to that in wild-type lymphocytes. In addition, we show that the scid mutation disturbs p460 nuclear association, presumably affecting its role in DNA repair pathways. To examine the molecular basis for our observations, we used a degenerate PCR strategy to clone the C-terminal p460 kinase domain from wild-type and scid thymocytes. Northern (RNA) analysis with these probes revealed normal steady-state p460 mRNA levels in scid cells, suggesting that the reduced abundance of p460 protein is due to a posttranscriptional defect. Sequence comparisons identified a single-base-pair alteration in the scid C-terminal p460 kinase domain, resulting in a premature stop codon. This mutation is predicted to truncate p460 by approximately 8 kDa, but it preserves the conserved motifs required for kinase activity in members of the phosphoinositidyl 3-kinase family. Despite a computed molecular weight alteration of less than 2%, we were able to visualize this difference by Western blot (immunoblot) analysis of wild-type and scid p460. These data demonstrate that the scid DNA-PKes mutation is not a null allele and suggest a molecular rationale for the well-described leakiness of the scid phenotype.