The p97 segregase cofactor Ubxn7 facilitates replisome disassembly during S-phase

Complex cellular processes are driven by the regulated assembly and disassembly of large multiprotein complexes. While we are beginning to understand the molecular mechanism for assembly of the eukaryotic DNA replication machinery (replisome), we still know relatively little about the regulation of its disassembly at replication termination. Recently, the first elements of this process have emerged, revealing that the replicative helicase, at the heart of the replisome, is polyubiquitylated prior to unloading and that this unloading requires p97 segregase activity. Two different E3 ubiquitin ligases have now been shown to ubiquitylate the helicase under different conditions: Cul2^Lrr1 and TRAIP. Here, using Xenopus laevis egg extract cell-free system and biochemical approaches, we have found two p97 cofactors, Ubxn7 and Faf1, which can interact with p97 during replisome disassembly during S-phase. We show only Ubxn7, however, facilitates efficient replisome disassembly. Ubxn7 delivers this role through its interaction via independent domains with both Cul2^Lrr1 and p97 to allow coupling between Mcm7 ubiquitylation and its removal from chromatin. Our data therefore characterize Ubxn7 as the first substrate-specific p97 cofactor regulating replisome disassembly in vertebrates and a rationale for the efficacy of the Cul2^Lrr1 replisome unloading pathway in unperturbed S-phase.     

Impact of patient setup error in the treatment of head and neck cancer with intensity modulated radiation therapy

PurposeTo study the impact of setup errors on the dose to the target volume and critical structures in the treatment of cancer of nasopharynx with intensity modulated radiation therapy (IMRT).Methods and materialsTwelve patients of carcinoma of nasopharynx treated by IMRT with simultaneous integrated boost technique were enrolled. The gross tumor volume, clinical target volume and low-risk nodal region were planned for 70, 59.4 and 54 Gy, respectively, in 33 fractions. Based on the constraints, treatment plans were generated. Keeping it as the base plan, the patient setup error was simulated for 3, 5 and 10 mm by shifting the isocenter in all three directions viz. anterior, posterior, superior, inferior, right and left lateral. The plans were evaluated for mean dose, maximum dose, volume of PTV receiving >110% and <93% of the prescribed dose. For both the parotids, the mean dose and the dose received by >50% of the parotid were evaluated. The maximum dose and dose received by 2 cc of spinal cord were also analyzed.ResultsThe dose to the target volume decreases gradually with increase in setup error. The superior and inferior shifts play major role in tumor under-dosage. A setup error of 3 mm along the posterior and lateral directions significantly affects the dose to the spinal cord. Similarly, setup error along lateral and anterior directions affects the dose to both parotids.ConclusionsThe isocenter position should be verified regularly to ensure that the goal of IMRT is achieved.     

Radioprotection by α-asarone: prevention of genotoxicity and hematopoietic injury in mammalian organism

α-Asarone (1-propenyl-2,4,5-methoxybenzol), one of the active components of Acorus calamus extract, was examined for its efficacy as a radioprotector in mice exposed to lethal and sublethal whole-body γ-radiation. Oral administration of α-asarone 1h prior to the radiation exposure reduced radiation induced alterations in the endogenous antioxidant defense systems. The radiation induced cellular DNA damages as revealed by comet assay, micronuclei formation and chromosomal aberrations were also significantly reduced following the asarone treatment. α-Asarone administration enhanced the endogenous spleen colony formation and reduced radiation-induced mortality and facilitated recovery from the radiation-induced loss of body weight in mice surviving after 8Gy γ-radiation exposure. These studies highlight the role of α-asarone as a good natural radioprotecting agent with therapeutic implications in case of radiation-exposure scenarios.