ALBERT

Description: Genomic integrity is compromised by DNA polymerase replication errors, which occur in a sequence-dependent manner across the genome. Accurate and complete quantification of a DNA polymerase's error spectrum is challenging because errors are rare and difficult to detect. We report a high-throughput sequencing assay to map in vitro DNA replication errors at the single-molecule level. Unlike previous methods, our assay is able to rapidly detect a large number of polymerase errors at base resolution over any template substrate without quantification bias. To overcome the high error rate of high-throughput sequencing, our assay uses a barcoding strategy in which each replication product is tagged with a unique nucleotide sequence before amplification. This allows multiple sequencing reads of the same product to be compared so that sequencing errors can be found and removed. We demonstrate the ability of our assay to characterize the average error rate, error hotspots and lesion bypass fidelity of several DNA polymerases.

Description: Etheno DNA adducts are a prevalent type of DNA damage caused by vinyl chloride (VC) exposure and oxidative stress. Etheno adducts are mutagenic and may contribute to the initiation of several pathologies; thus, elucidating the pathways by which they induce cellular transformation is critical. Although N 2 ,3-ethenoguanine ( N 2 ,3-G) is the most abundant etheno adduct, its biological consequences have not been well characterized in cells due to its labile glycosidic bond. Here, a stabilized 2'-fluoro-2'-deoxyribose analog of N 2 ,3-G was used to quantify directly its genotoxicity and mutagenicity. A multiplex method involving next-generation sequencing enabled a large-scale in vivo analysis, in which both N 2 ,3-G and its isomer 1, N 2 -ethenoguanine (1, N 2 -G) were evaluated in various repair and replication backgrounds. We found that N 2 ,3-G potently induces G to A transitions, the same mutation previously observed in VC-associated tumors. By contrast, 1, N 2 -G induces various substitutions and frameshifts. We also found that N 2 ,3-G is the only etheno lesion that cannot be repaired by AlkB, which partially explains its persistence. Both G lesions are strong replication blocks and DinB, a translesion polymerase, facilitates the mutagenic bypass of both lesions. Collectively, our results indicate that N 2 ,3-G is a biologically important lesion and may have a functional role in VC-induced or inflammation-driven carcinogenesis.

Description: Seismic ambient noise tomography is applied to central and southern Mozambique, located in the tip of the East African Rift (EAR). The deployment of MOZART seismic network, with a total of 30 broad-band stations continuously recording for 26 months, allowed us to carry out the first tomographic study of the crust under this region, which until now remained largely unexplored at this scale. From cross-correlations extracted from coherent noise we obtained Rayleigh wave group velocity dispersion curves for the period range 5–40 s. These dispersion relations were inverted to produce group velocity maps, and 1-D shear wave velocity profiles at selected points. High group velocities are observed at all periods on the eastern edge of the Kaapvaal and Zimbabwe cratons, in agreement with the findings of previous studies. Further east, a pronounced slow anomaly is observed in central and southern Mozambique, where the rifting between southern Africa and Antarctica created a passive margin in the Mesozoic, and further rifting is currently happening as a result of the southward propagation of the EAR. In this study, we also addressed the question concerning the nature of the crust (continental versus oceanic) in the Mozambique Coastal Plains (MCP), still in debate. Our data do not support previous suggestions that the MCP are floored by oceanic crust since a shallow Moho could not be detected, and we discuss an alternative explanation for its ocean-like magnetic signature. Our velocity maps suggest that the crystalline basement of the Zimbabwe craton may extend further east well into Mozambique underneath the sediment cover, contrary to what is usually assumed, while further south the Kaapval craton passes into slow rifted crust at the Lebombo monocline as expected. The sharp passage from fast crust to slow crust on the northern part of the study area coincides with the seismically active NNE-SSW Urema rift, while further south the Mazenga graben adopts an N-S direction parallel to the eastern limit of the Kaapvaal craton. We conclude that these two extensional structures herald the southward continuation of the EAR, and infer a structural control of the transition between the two types of crust on the ongoing deformation.

Description: Escherichia coli has three DNA polymerases implicated in the bypass of DNA damage, a process called translesion synthesis (TLS) that alleviates replication stalling. Although these polymerases are specialized for different DNA lesions, it is unclear if they interact differently with the replication machinery. Of the three, DNA polymerase (Pol) II remains the most enigmatic. Here we report a stable ternary complex of Pol II, the replicative polymerase Pol III core complex and the dimeric processivity clamp, β. Single-molecule experiments reveal that the interactions of Pol II and Pol III with β allow for rapid exchange during DNA synthesis. As with another TLS polymerase, Pol IV, increasing concentrations of Pol II displace the Pol III core during DNA synthesis in a minimal reconstitution of primer extension. However, in contrast to Pol IV, Pol II is inefficient at disrupting rolling-circle synthesis by the fully reconstituted Pol III replisome. Together, these data suggest a β-mediated mechanism of exchange between Pol II and Pol III that occurs outside the replication fork.

Description: Antedependence models, also known as transition models, have proven to be useful for longitudinal data exhibiting serial correlation, especially when the variances and/or same-lag correlations are time-varying. Statistical inference procedures associated with normal antedependence models are well-developed and have many nice properties, but they are not appropriate for longitudinal data that exhibit considerable skewness. We propose two direct extensions of normal antedependence models to skew-normal antedependence models. The first is obtained by imposing antedependence on a multivariate skew-normal distribution, and the second is a sequential autoregressive model with skew-normal innovations. For both models, necessary and sufficient conditions for $p$ th-order antedependence are established, and likelihood-based estimation and testing procedures for models satisfying those conditions are developed. The procedures are applied to simulated data and to real data from a study of cattle growth.

Description: The unification scheme of active galactic nuclei invokes an optically thick molecular torus component hiding the broad emission line region. Assuming the presence of a thick neutral component in the molecular torus characterized by a H  i column density 〉10 22 cm –2 , we propose that far-UV radiation around Lyα can be significantly polarized through Rayleigh scattering. Adopting a Monte Carlo technique, we compute polarization of Rayleigh scattered radiation near Lyα in a thick neutral region in the shape of a slab and a cylindrical shell. It is found that radiation near Lyα Rayleigh reflected from a very thick slab can be significantly polarized in a fairly large range of wavelength  ~ 50 Å exhibiting a flux profile similar to the incident one. Rayleigh transmitted radiation in a slab is characterized by the central dip with a complicated polarization behaviour. The optically thick part near Lyα centre is polarized in the direction perpendicular to the slab normal, which is in contrast to weakly polarized wing parts in the direction parallel to the slab normal. A similar polarization flip phenomenon is also found in the case of a tall cylindrical shell, in which the spatial diffusion along the vertical direction near the inner cylinder wall for core photons leads to a tendency of the electric field aligned to the direction perpendicular to the vertical axis. Observational implications are briefly discussed including spectropolarimetry of the quasar PG 1630+377 by Koratkar et al. in 1990 where Lyα is strongly polarized with no other emission lines polarized.

Description: Three gamma-ray millisecond pulsars (MSPs), PSR J1939+2134, PSR J1959+2048, and PSR J0034-0534, have been confirmed to have a common feature of phase-aligned in radio and gamma-ray bands. With a geometric (two-pole caustic) model and a physical outer gap model (revised 3D outer gap model) in a three dimensional (3D) retarded magnetic dipole with a perturbation magnetic field, the observed features of these MSPs are studied. In order to obtained the best-fitting model parameters, the Markov chain Monte Carlo technique is used and reasonable GeV band light curves for three MSPs are given. Our calculations indicate that MSPs emit high energy photons with smaller inclination angles (α ≈ 10°–50°), larger view angles (ζ ≈ 65°–100°), and smaller perturbation factor (ε ≈ −0.15–0.1). Note that the factor ε, describing the strength of the perturbed magnetic field, is all less than zero in these two models, so the magnetic field caused by current-induced play a leading role in the pulsed location of MSPs.

Description: We previously presented the YM500 database, which contains 〉8000 small RNA sequencing (smRNA-seq) data sets and integrated analysis results for various cancer miRNome studies. In the updated YM500v3 database ( http://ngs.ym.edu.tw/ym500/ ) presented herein, we not only focus on miRNAs but also on other functional small non-coding RNAs (sncRNAs), such as PIWI-interacting RNAs (piRNAs), tRNA-derived fragments (tRFs), small nuclear RNAs (snRNAs) and small nucleolar RNAs (snoRNAs). There is growing knowledge of the role of sncRNAs in gene regulation and tumorigenesis. We have also incorporated 〉10 000 cancer-related RNA-seq and 〉3000 more smRNA-seq data sets into the YM500v3 database. Furthermore, there are two main new sections, ‘Survival' and ‘Cancer', in this updated version. The ‘Survival’ section provides the survival analysis results in all cancer types or in a user-defined group of samples for a specific sncRNA. The ‘Cancer’ section provides the results of differential expression analyses, miRNA–gene interactions and cancer miRNA-related pathways. In the ‘Expression’ section, sncRNA expression profiles across cancer and sample types are newly provided. Cancer-related sncRNAs hold potential for both biotech applications and basic research.