Recent advances in the structural mechanisms of DNA glycosylases

DNA glycosylases safeguard the genome by locating and excising a diverse array of aberrant nucleobases created from oxidation, alkylation, and deamination of DNA. Since the discovery 28 years ago that these enzymes employ a base flipping mechanism to trap their substrates, six different protein architectures have been identified to perform the same basic task. Work over the past several years has unraveled details for how the various DNA glycosylases survey DNA, detect damage within the duplex, select for the correct modification, and catalyze base excision. Here, we provide a broad overview of these latest advances in glycosylase mechanisms gleaned from structural enzymology, highlighting features common to all glycosylases as well as key differences that define their particular substrate specificities.     

Invaded range of the blackberry pathogen Phragmidium violaceum in the Pacific Northwest of the USA and the search for its provenance

Field surveys in 2006 confirmed that the exotic rust fungus Phragmidium violaceum was widespread on Rubus armeniacus and Rubus laciniatus in the Pacific Northwest of the USA. The origin and dispersal pattern of this obligate biotrophic pathogen in the USA were investigated by comparing the genetic diversity and structure of 27 isolates each from the USA and Europe, and 20 isolates from Australia where an invasion occurred in 1984. Analysis of 11 microsatellite loci revealed 74 unique genotypes, with the European population having a significantly higher level of allelic diversity and number of private alleles compared to populations from the USA and Australia. Principal coordinate analysis (PCA), analysis of molecular variance and pairwise comparisons of Φ confirmed a strong level of differentiation among continental populations, with little divergence between isolates from the USA and Europe, but a high level of differentiation between these isolates and those from Australia. These results were broadly supported by the Bayesian cluster analysis, which indicated that at K = 3 the clustering of the isolates corresponds to their geographic origin. Bayesian clustering, PCA as well as insignificant migration estimates from Europe to the USA suggest that the USA population is not a direct descendant from the European P. violaceum population. There was a weak association between genetic and geographic distance among the USA isolates, suggesting invasion was initially localized prior to dispersal or that the population may have been present for some time prior to first detection in 2005.     

Growth of Greater White-Fronted Goose Goslings Relates to Population Dynamics at Multiple Scales

The abundance of greater white-fronted geese (Anser albifrons frontalis) on the Arctic Coastal Plain (ACP) of northern Alaska, USA, has more than tripled since the late 1990s; however, recent rate of annual population growth has declined as population size increased, which may indicate white-fronted geese on the ACP are approaching carrying capacity. We examined rates of gosling growth in greater white-fronted geese at 3 sites on the ACP during 2012–2014 to assist with predictions of future population trends and assess evidence for density-dependent constraints on recruitment. We marked goslings at hatch with individually coded webtags and conducted brood drives during early August to capture, measure, and weigh goslings. Annual estimates of gosling mass at 32 days old (range = 1,190–1,685) indicate that goslings had obtained >60% of asymptotic size. This rate of growth corresponds with that of other goose species and populations with access to high-quality forage and no limitations on forage availability, and is consistent with the overall increase in abundance of white-fronted geese at the ACP scale. Contrary to most previous investigations, age-adjusted mass of goslings did not decline with hatch date. Goslings grew faster in coastal areas than at inland freshwater sites. Taken together, these findings suggest forage was not limiting gosling growth rates in either ecosystem, but forage was of greater quality in coastal areas where goose foraging habitat is expanding because of permafrost subsidence. Spatial patterns of gosling growth corresponded with local-scale patterns of population density and population change; the areas with greatest rates of gosling growth were those with the greatest population density and rates of population increase. We found little evidence to suggest forage during brood rearing was limiting population increase of white-fronted geese on the ACP. Factors responsible for the apparent slowing of ACP-wide population growth are likely those that occur in stages of the annual cycle outside of the breeding grounds. Published 2021. This article is a U.S. Government work and is in the public domain in the USA.     

RSEQtools: a modular framework to analyze RNA-Seq data using compact, anonymized data summaries

SUMMARY: The advent of next-generation sequencing for functional genomics has given rise to quantities of sequence information that are often so large that they are difficult to handle. Moreover, sequence reads from a specific individual can contain sufficient information to potentially identify and genetically characterize that person, raising privacy concerns. In order to address these issues, we have developed the Mapped Read Format (MRF), a compact data summary format for both short and long read alignments that enables the anonymization of confidential sequence information, while allowing one to still carry out many functional genomics studies. We have developed a suite of tools (RSEQtools) that use this format for the analysis of RNA-Seq experiments. These tools consist of a set of modules that perform common tasks such as calculating gene expression values, generating signal tracks of mapped reads and segmenting that signal into actively transcribed regions. Moreover, the tools can readily be used to build customizable RNA-Seq workflows. In addition to the anonymization afforded by MRF, this format also facilitates the decoupling of the alignment of reads from downstream analyses. Availability and implementation: RSEQtools is implemented in C and the source code is available at http://rseqtools.gersteinlab.org/.     

Sun-dried raisins are a cost-effective alternative to Sports Jelly Beans in prolonged cycling

The purpose of this study was to examine the effects of a natural carbohydrate (CHO) source in the form of sun-dried raisins (SDRs) vs. Sports Jelly Beans? (SJBs) on endurance performance in trained cyclists and triathletes. Ten healthy men (18-33 years) completed 1 water-only acclimatization exercise trial and 2 randomized exercise trials administered in a crossover fashion. Each trial consisted of a 120-minute constant-intensity glycogen depletion period followed by a 10-km time trial (TT). During each experimental trial, participants consumed isocaloric amounts of SDRs or SJBs in 20-minute intervals. Measurements included time to complete 10-km TT, power output during 10-km TT, blood glucose levels and respiratory exchange ratio during glycogen depletion period, rate of perceived exertion (RPE), 'flow' questionnaire responses, and a hedonic (i.e., pleasantness) sensory acceptance test. There were no significant differences in endurance performance for TT time (SDRs vs. SJBs, 17.3 ± 0.4 vs. 17.3 ± 0.4 seconds) or power (229.3 ± 13.0 vs. 232.0 ± 13.6 W), resting blood glucose levels (5.8 ± 04 mmol·L(-1) for SDRs and 5.4 ± 0.2 mmol·L(-1) for SJBs), RPE, or flow experiences between SDR and SJB trials. However, the mean sensory acceptance scores were significantly higher for the SDRs compared to the SJBs (50.7 ± 1.7 vs. 44.3 ± 2.7). Consuming SDRs or SJBs during 120 minutes of intense cycling results in similar subsequent TT performances and are equally effective in maintaining blood glucose levels during exercise. Therefore, SDRs are a natural, pleasant, cost-effective CHO alternative to commercial SJBs that can be used during moderate- to high-intensity endurance exercise.     

Unraveling a connection between DNA demethylation repair and cancer

In this issue of Molecular Cell, Dango and Mosammaparast discover that the human oxidative demethylase ALKBH3 functions in complex with a DNA helicase to eliminate N3-methylcytosine lesions from ssDNA and that specific cancer cell lines are dependent on this activity for proliferation (Dango et al., 2011).     

Informed consent and genomic incidental findings: IRB chair perspectives

It is unclear how genomic incidental finding (GIF) prospects should be addressed in informed consent processes. An exploratory study on this topic was conducted with 34 purposively sampled Chairs of institutional review boards (IRBs) at centers conducting genome-wide association studies. Most Chairs (96%) reported no knowledge of local IRB requirements regarding GIFs and informed consent. Chairs suggested consent processes should address the prospect of, and study disclosure policy on, GIFs; GIF management and follow-up; potential clinical significance of GIFs; potential risks of GIF disclosure; an opportunity for participants to opt out of GIF disclosure; and duration of the researcher's duty to disclose GIFs. Chairs were concerned about participant disclosure preferences changing over time; inherent limitations in determining the scope and accuracy of claims about GIFs; and making consent processes longer and more complex. IRB Chair and other stakeholder perspectives can help advance informed consent efforts to accommodate GIF prospects.