Biology subjects

Chase, J.

Publications and source records attributed to Chase, J..

21 records · Page 2Linked to original sources

bioRxiv2017

Routine Single Particle CryoEM Sample and Grid Characterization by Tomography

Single particle cryo-electron microscopy (cryoEM) is often performed under the assumption that particles are freely floating away from the air-water interfaces and in thin, vitreous ice. In this study, we performed fiducial-less tomography on over 50 different cryoEM grid/sample preparations to determine the particle distribution within the ice and the overall geometry of the ice in grid holes. Surprisingly, by studying particles in holes in 3D from over 1,000 tomograms, we have determined that the vast majority of particles (approximately 90%) are adsorbed to an air-water interface. The implications of this observation are wide-ranging, with potential ramifications regarding protein denaturation, conformational change, and preferred orientation. We also show that fiducial-less cryo-electron tomography on single particle grids may be used to determine ice thickness, optimal single particle collection areas and strategies, particle heterogeneity, and de novo models for template picking and single particle alignment.

biophysics↗

bioRxiv2017

Copy number heterogeneity, large origin tandem repeats, and interspecies recombination in HHV-6A and HHV-6B reference strains

Quantitative PCR is the diagnostic pillar for clinical virology testing, and reference materials are necessary for accurate, comparable quantitation between clinical laboratories. Accurate quantitation of HHV-6 is important for detection of viral reactivation and inherited chromosomally integrated HHV-6 in immunocompromised patients. Reference materials in clinical virology commonly consist of laboratory-adapted viral strains that may be affected by the culture process. We performed next-generation sequencing to make relative copy number measurements at single nucleotide resolution of eight candidate HHV-6A and seven HHV-6B reference strains and DNA materials from the HHV-6 Foundation and Advanced Biotechnologies. 11 of 17 (65%) HHV6 candidate reference materials showed multiple copies of the origin of replication upstream of the U41 gene by next-generation sequencing. These large tandem repeats arose independently in culture-adapted HHV-6A and HHV-6B strains, measuring 1254 bp and 983 bp, respectively. Copy number measured between 4-10X copies relative to the rest of the genome. We also report the first interspecies recombinant HHV-6 strain with a HHV-6A GS backbone and >5.5kb region from HHV-6B Z29 from U41-U43 that covered the origin tandem repeat. Specific HHV-6A reference strains demonstrated duplication of regions at UL1/UL2, U87, and U89, as well as deletion in the U12-U24 region and U94/95 genes. HHV-6 strains derived from cord blood mononuclear cells from different labs on different continents revealed no copy number differences throughout the viral genome. These data indicate large origin tandem duplications are an adaptation of both HHV-6A and HHV-6B in culture and show interspecies recombination is possible within the Betaherpesvirinae.\n\nImportanceAnything in science that needs to be quantitated requires a standard unit of measurement. This includes viruses, for which quantitation increasingly determines definitions of pathology and guidelines for treatment. However, the act of making standard or reference material in virology can alter its very usefulness through genomic duplications, insertions, and rearrangements. We used deep sequencing to examine candidate reference strains for HHV-6, a ubiquitous human virus that can reactivate in the immunocompromised population and is integrated into the human genome in every cell of the body for 1% of people worldwide. We found large tandem repeats in the origin of replication for both HHV-6A and HHV-6B that are selected for in culture. We also found the first interspecies recombinant between HHV-6A and HHV-6B, a phenomenon that is well-known in alphaherpesviruses but to date has not been seen in betaherpesviruses. These data critically inform HHV-6 biology and the standard selection process.

genomics↗

bioRxiv2016

Extinction rate has a complex and non-linear relationship with area

Aim.Biodiversity loss, measured as count of extinction events, is a key component of biodiversity change, and can significantly impact ecosystem services. However, estimation of the loss has focused mostly on per-species extinction rates measured over limited numbers of spatial scales, with no theory linking small-scale extirpations with global extinctions. Here we provide such link by introducing the relationship between area and per-species probability of extinction (PxAR) and between area and count of realized extinction events in that area (NxAR). We show theoretical and empirical forms of these relationships, and we discuss their role in perception and estimation of the current extinction crisis.\n\nLocationUSA, Europe, Czech Republic, Barro Colorado Island\n\nMethodsWe derived the expected forms of PxAR and NxAR from a range of theoretical frameworks based on theory of island biogeography, neutral models, and species-area relationships. We constructed PxAR and NxAR in five empirical datasets on butterflies, plants, trees and birds, collected over range of spatial scales.\n\nResultsBoth the theoretical arguments and empirical data support monotonically decreasing PxAR, i.e. per-species extinction probability decreasing with increasing area; however, we also report a rare theoretical possibility of locally increasing PxAR. In contrast, both theory and data revealed complex NxAR, i.e. counts of extinction events follow variety of relationships with area, including nonlinear unimodal, multimodal and U-shaped relationships, depending on region and taxon.\n\nMain conclusionsThe uncovered wealth of forms of NxAR can explain why biodiversity change (the net outcome of losses and gains) also appears scale-dependent. Furthermore, the complex scale dependence of PxAR and NxAR means that global extinctions indicate little about local extirpations, and vice versa. Hence, effort should be made to understand and report extinction crisis as a scale-dependent problem. In this effort, estimation of scaling relationships such as PxAR and NxAR should be central.

ecology↗