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Biology subjects

Kelly, J. W.

Publications and source records attributed to Kelly, J. W..

4 recordsLinked to original sources

Quantitative Interactome Proteomics Reveals a Molecular Basis for ATF6-Dependent Regulation of a Destabilized Amyloidogenic Protein

Activation of the unfolded protein response (UPR)-associated transcription factor ATF6 has emerged as a promising strategy to selectively reduce the secretion and subsequent toxic aggregation of destabilized, amyloidogenic proteins implicated in diverse systemic amyloid diseases. However, the molecular mechanism by which ATF6 activation reduces the secretion of amyloidogenic proteins remains poorly defined. Here, we establish a quantitative interactomics platform with improved throughput and sensitivity to define how ATF6 activation selectively reduces secretion of a destabilized, amyloidogenic immunoglobulin light chain (LC) associated with Light Chain Amyloidosis (AL). We show that ATF6 activation increases the targeting of this destabilized LC to a select subset of pro-folding ER proteostasis factors that retains the amyloidogenic LC within the ER, preventing its secretion to downstream secretory environments. Our results define a molecular basis for the selective, ATF6-dependent reduction in destabilized LC secretion and highlight the advantage for targeting this endogenous UPR-associated transcription factor to reduce secretion of destabilized, amyloidogenic proteins implicated in AL and related systemic amyloid diseases.

cell biology

Crowdsourcing Image Analysis for Plant Phenomics to Generate Ground Truth Data for Machine Learning

The accuracy of machine learning tasks critically depends on high quality ground truth data. Therefore, in many cases, producing good ground truth data typically involves trained professionals; however, this can be costly in time, effort, and money. Here we explore the use of crowdsourcing to generate a large number of training data of good quality. We explore an image analysis task involving the segmentation of corn tassels from images taken in a field setting. We investigate the accuracy, speed and other quality metrics when this task is performed by students for academic credit, Amazon MTurk workers, and Master Amazon MTurk workers. We conclude that the Amazon MTurk and Master Mturk workers perform significantly better than the for-credit students, but with no significant difference between the two MTurk worker types. Furthermore, the quality of the segmentation produced by Amazon MTurk workers rivals that of an expert worker. We provide best practices to assess the quality of ground truth data, and to compare data quality produced by different sources. We conclude that properly managed crowdsourcing can be used to establish large volumes of viable ground truth data at a low cost and high quality, especially in the context of high throughput plant phenotyping. We also provide several metrics for assessing the quality of the generated datasets.\n\nAuthor SummaryFood security is a growing global concern. Farmers, plant breeders, and geneticists are hastening to address the challenges presented to agriculture by climate change, dwindling arable land, and population growth. Scientists in the field of plant phenomics are using satellite and drone images to understand how crops respond to a changing environment and to combine genetics and environmental measures to maximize crop growth efficiency. However, the terabytes of image data require new computational methods to extract useful information. Machine learning algorithms are effective in recognizing select parts of images, but they require high quality data curated by people to train them, a process that can be laborious and costly. We examined how well crowdsourcing works in providing training data for plant phenomics, specifically, segmenting a corn tassel - the male flower of the corn plant - from the often-cluttered images of a cornfield. We provided images to students, and to Amazon MTurkers, the latter being an on-demand workforce brokered by Amazon.com and paid on a task-by-task basis. We report on best practices in crowdsourcing image labeling for phenomics, and compare the different groups on measures such as fatigue and accuracy over time. We find that crowdsourcing is a good way of generating quality labeled data, rivaling that of experts.

plant biology

Amyloid accumulation drives proteome-wide alterations in mouse models of Alzheimers disease like pathology

Amyloid beta (A{beta}) peptides impair multiple cellular pathways in the brain and play a causative role in Alzheimers disease (AD) pathology, but how the brain proteome is remodeled during this process is unknown. To identify new protein networks associated with AD-like pathology, we performed global quantitative proteomic analysis in three mouse models at pre- and post-symptomatic ages. Our analysis revealed a robust and consistent increase in Apolipoprotein E (ApoE) levels in nearly all transgenic brain regions with increased A{beta} levels. Taken together with prior findings on ApoE driving A{beta} accumulation, this analysis points to a pathological dysregulation of the ApoE-A{beta} axis. We also found dysregulation of protein networks involved in excitatory synaptic transmission consistent with AD pathophysiology. Targeted analysis of the AMPA receptor complex revealed a specific loss of TARP{gamma}-2, a key AMPA receptor trafficking protein. Expression of TARP{gamma}-2 in vivo in hAPP transgenic mice led to a restoration of AMPA currents. This database of proteome alterations represents a unique resource for the identification of protein alterations responsible for AD.\n\nHighlightsO_LIProteomic analysis of mouse brains with AD-like pathology reveals stark remodeling\nC_LIO_LIProteomic evidence points to a dysregulation of ApoE levels associated with A{beta} clearance rather than production\nC_LIO_LICo-expression analysis found distinctly impaired synapse and mitochondria modules\nC_LIO_LIIn-depth analyses of AMPAR complex points to loss of TARP{gamma}-2, which may compromise synapses in AD\nC_LI\n\neTOC BlurbProteome-wide profiling of brain tissue from three mouse models of AD-like pathology reveals A{beta}, brain region, and age dependent alterations of protein levels. This resource provides a new global protein expression atlas for the Alzheimers disease research community.

neuroscience

Translation efficiency is maintained at elevated temperature in E. coli

Cellular protein levels are dictated by the balance between gene transcription, mRNA translation and protein degradation, among other factors. Cells must manage their proteomes during stress; one way in which they may do so, in principle, is by differential translation. We used ribosome profiling to directly monitor translation in E. coli at 30 {degrees}C and investigate how this changes after 10-20 minutes of heat shock at 42 {degrees}C. Translation is controlled by the interplay of several RNA hybridization processes, which are expected to be temperature sensitive. However, translation efficiencies are robustly maintained after thermal heat shock and after mimicking the heat shock response transcriptional program at 30 {degrees}C. Several gene-specific parameters correlated with translation efficiency, including predicted mRNA structure and whether a gene is cotranslationally translocated into the inner membrane. Genome-wide predictions of the temperature dependence of mRNA structure suggest that relatively few genes show a melting transition between 30 {degrees}C and 42 {degrees}C, consistent with our observations. A linear model with five parameters can predict 33% of the variation in translation efficiency between genes, which may be useful in interpreting transcriptome data.

biochemistry