BCCIP regulates homologous recombination by distinct domains and suppresses spontaneous DNA damage

Homologous recombination (HR) is critical for maintaining genome stability through precise repair of DNA double-strand breaks (DSBs) and restarting stalled or collapsed DNA replication forks. HR is regulated by many proteins through distinct mechanisms. Some proteins have direct enzymatic roles in HR reactions, while others act as accessory factors that regulate HR enzymatic activity or coordinate HR with other cellular processes such as the cell cycle. The breast cancer susceptibility gene BRCA2 encodes a critical accessory protein that interacts with the RAD51 recombinase and this interaction fluctuates during the cell cycle. We previously showed that a BRCA2- and p21-interacting protein, BCCIP, regulates BRCA2 and RAD51 nuclear focus formation, DSB-induced HR and cell cycle progression. However, it has not been clear whether BCCIP acts exclusively through BRCA2 to regulate HR and whether BCCIP also regulates the alternative DSB repair pathway, non-homologous end joining. In this study, we found that BCCIP fragments that interact with BRCA2 or with p21 each inhibit DSB repair by HR. We further show that transient down-regulation of BCCIP in human cells does not affect non-specific integration of transfected DNA, but significantly inhibits homology-directed gene targeting. Furthermore, human HT1080 cells with constitutive down-regulation of BCCIP display increased levels of spontaneous single-stranded DNA (ssDNA) and DSBs. These data indicate that multiple BCCIP domains are important for HR regulation, that BCCIP is unlikely to regulate non-homologous end joining, and that BCCIP plays a critical role in resolving spontaneous DNA damage

Requirement of Mouse BCCIP for Neural Development and Progenitor Proliferation

Multiple DNA repair pathways are involved in the orderly development of neural systems at distinct stages. The homologous recombination (HR) pathway is required to resolve stalled replication forks and critical for the proliferation of progenitor cells during neural development. BCCIP is a BRCA2 and CDKN1A interacting protein implicated in HR and inhibition of DNA replication stress. In this study, we determined the role of BCCIP in neural development using a conditional BCCIP knock-down mouse model. BCCIP deficiency impaired embryonic and postnatal neural development, causing severe ataxia, cerebral and cerebellar defects, and microcephaly. These development defects are associated with spontaneous DNA damage and subsequent cell death in the proliferative cell populations of the neural system during embryogenesis. With in vitro neural spheroid cultures, BCCIP deficiency impaired neural progenitor's self-renewal capability, and spontaneously activated p53. These data suggest that BCCIP and its anti-replication stress functions are essential for normal neural development by maintaining an orderly proliferation of neural progenitors

Essential Roles of BCCIP in Mouse Embryonic Development and Structural Stability of Chromosomes

BCCIP is a BRCA2- and CDKN1A(p21)-interacting protein that has been implicated in the maintenance of genomic integrity. To understand the in vivo functions of BCCIP, we generated a conditional BCCIP knockdown transgenic mouse model using Cre-LoxP mediated RNA interference. The BCCIP knockdown embryos displayed impaired cellular proliferation and apoptosis at day E7.5. Consistent with these results, the in vitro proliferation of blastocysts and mouse embryonic fibroblasts (MEFs) of BCCIP knockdown mice were impaired considerably. The BCCIP deficient mouse embryos die before E11.5 day. Deletion of the p53 gene could not rescue the embryonic lethality due to BCCIP deficiency, but partially rescues the growth delay of mouse embryonic fibroblasts in vitro. To further understand the cause of development and proliferation defects in BCCIP-deficient mice, MEFs were subjected to chromosome stability analysis. The BCCIP-deficient MEFs displayed significant spontaneous chromosome structural alterations associated with replication stress, including a 3.5-fold induction of chromatid breaks. Remarkably, the BCCIP-deficient MEFs had a ∼20-fold increase in sister chromatid union (SCU), yet the induction of sister chromatid exchanges (SCE) was modestly at 1.5 fold. SCU is a unique type of chromatid aberration that may give rise to chromatin bridges between daughter nuclei in anaphase. In addition, the BCCIP-deficient MEFs have reduced repair of irradiation-induced DNA damage and reductions of Rad51 protein and nuclear foci. Our data suggest a unique function of BCCIP, not only in repair of DNA damage, but also in resolving stalled replication forks and prevention of replication stress. In addition, BCCIP deficiency causes excessive spontaneous chromatin bridges via the formation of SCU, which can subsequently impair chromosome segregations in mitosis and cell division

Alterations of BCCIP, a BRCA2 interacting protein, in astrocytomas

<p>Abstract</p> <p>Background</p> <p>Loss of heterozygosity of chromosome 10q26 has been shown to be associated with the aggressiveness of astrocytic tumors (or astrocytomas), but the responsible gene(s) residing in this region has not been fully identified. The <it>BCCIP </it>gene is located at chromosome 10q26. It encodes a BRCA2 and CDKN1A (p21) interacting protein. Previous studies have shown that down-regulation of BCCIP impairs recombinational DNA repair, G1/S cell cycle checkpoint, p53 trans-activation activity, cytokinesis, and chromosome stability, suggesting a potential role of <it>BCCIP </it>in cancer etiology. In this study, we investigated whether <it>BCCIP </it>is altered in astrocytomas.</p> <p>Methods</p> <p>Genomic DNA from 45 cases of grade IV astrocytic tumor (glioblastoma) tissues and 12 cases of normal tissues were analyzed by quantitative PCR. The BCCIP protein expression in 96 cases of grade II–IV astrocytic tumors was detected by immunohistochemistry (IHC). IHC staining of glial fibrillary acid protein (GFAP), a marker for astrocytic cells, was used to identify cells of the astrocytic lineage.</p> <p>Results</p> <p>We found that BCCIP protein is expressed in normal cells with positive staining of GFAP. However, BCCIP protein expression was not detectable in ~45% of all astrocytic tumors, and in > 60% in the grade IV glioblastoma. About 45% glioblastoma have significant (p < 0.01) reduction of <it>BCCIP </it>gene copy number when compared to normal DNA. Furthermore, the frequency of lacking BCCIP expression is associated with the aggressiveness of astrocytic tumors.</p> <p>Conclusion</p> <p>Our data implicate a role of BCCIP in astrocytic tumorigenesis, and lack of <it>BCCIP </it>may be used as a marker for astrocytomas.</p

Almost Periodic Synchronization of Fuzzy Cellular Neural Networks with Time–Varying Delays via State–Feedback and Impulsive Control

In this paper, we are concerned with drive-response synchronization for a class of fuzzy cellular neural networks with time varying delays. Based on the exponential dichotomy of linear differential equations, the Banach fixed point theorem and the differential inequality technique, we obtain the existence of almost periodic solutions of this class of networks. Then, we design a state feedback and an impulsive controller, and construct a suitable Lyapunov function to study the problem of global exponential almost periodic synchronization for the drive-response systems considered. At the end of the paper, we provide an example to verify the effectiveness of the theoretical results

Almost Periodic Synchronization of Fuzzy Cellular Neural Networks with Time–Varying Delays via State–Feedback and Impulsive Control

In this paper, we are concerned with drive-response synchronization for a class of fuzzy cellular neural networks with time varying delays. Based on the exponential dichotomy of linear differential equations, the Banach fixed point theorem and the differential inequality technique, we obtain the existence of almost periodic solutions of this class of networks. Then, we design a state feedback and an impulsive controller, and construct a suitable Lyapunov function to study the problem of global exponential almost periodic synchronization for the drive-response systems considered. At the end of the paper, we provide an example to verify the effectiveness of the theoretical results

Gender differences in thermal comfort in a hot-humid climate

Thirty female and 30 male college students, who had naturally acclimatized to hot-humid climate, were exposed to seven temperature/humidity combinations in a climate chamber, and they were surveyed all year long at their classrooms and dormitories. They dressed in 0.5 clo in the chamber tests, while their subjective and physiological responses were collected. In the field study the subjects were surveyed while the thermal parameters around their body were measured. Results show no significant differences in thermal sensitivity to temperature changes or in neutral temperatures, based on our chamber and field studies with the same group of subjects. Both genders were equally satisfied with temperatures, although females tended to be more dissatisfied in cool temperatures, and males tended to be more dissatisfied in warm temperatures in the chamber tests; however the differences disappeared when they can adjust their clothing in the field.

Aboveground Biomass in China’s Managed Grasslands and Their Responses to Environmental and Management Variations

Aboveground biomass (AGB) in managed grasslands can vary across a suite of environmental and management conditions; however, there lacks a quantitative assessment at the national scale of China. Although the potential effects of individual drivers (e.g., species, nutrient fertilization, and water management) have been examined in China’s managed grasslands, no attempts have been made to comprehensively assess the effects of multiple variables on AGB. Using a meta-data analysis approach, we created a database composed of AGB and associated attributes of managed grasslands in China. The database was used to assess the responses of AGB to anthropogenic factors, in addition to a suite of natural variables including climate, soil, and topography. The average AGB in managed grasslands of China is approximately 630 g m−2 of dry matter, ranging from 55 to 2172 g m−2 (95% confidence interval). Medicago sativa is the most widely planted species in China’s managed grasslands, followed by Elymus dahuricus and Bromus japonicus. The national average AGB of these three species was around 692, 530, and 856 g m−2, respectively. For each species, AGB shows a large discrepancy across different places. In general, grassland AGB depends substantially on species, environments, and management practices. The dependence can be well described by a linear mixed-effects regression in which a series of biotic and abiotic factors are used as predictors. We highlight that establishing managed grassland can potentially contribute to not only AGB enhancement, but also grassland restoration on degraded natural grasslands

Gender differences in thermal comfort in a hot-humid climate

Thirty female and 30 male college students, who had naturally acclimatized to hot-humid climate, were exposed to seven temperature/humidity combinations in a climate chamber, and they were surveyed all year long at their classrooms and dormitories. They dressed in 0.5 clo in the chamber tests, while their subjective and physiological responses were collected. In the field study the subjects were surveyed while the thermal parameters around their body were measured. Results show no significant differences in thermal sensitivity to temperature changes or in neutral temperatures, based on our chamber and field studies with the same group of subjects. Both genders were equally satisfied with temperatures, although females tended to be more dissatisfied in cool temperatures, and males tended to be more dissatisfied in warm temperatures in the chamber tests; however the differences disappeared when they can adjust their clothing in the field.