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Full-Text Articles in Life Sciences
Multi-Scale 3d Cryo-Correlative Microscopy For Vitrified Cells, Gong-Her Wu, Patrick G. Mitchell, Jesus G. Galaz-Montoya, Corey W. Hecksel, Emily M. Sontag, Vimal Gangadharan, Jeffrey Marshman, David Mankus, Margaret E. Bisher, Abigail K.R. Lytton-Jean, Judith Frydman, Kirk Czymmek, Wah Chiu
Multi-Scale 3d Cryo-Correlative Microscopy For Vitrified Cells, Gong-Her Wu, Patrick G. Mitchell, Jesus G. Galaz-Montoya, Corey W. Hecksel, Emily M. Sontag, Vimal Gangadharan, Jeffrey Marshman, David Mankus, Margaret E. Bisher, Abigail K.R. Lytton-Jean, Judith Frydman, Kirk Czymmek, Wah Chiu
Biological Sciences Faculty Research and Publications
Three-dimensional (3D) visualization of vitrified cells can uncover structures of subcellular complexes without chemical fixation or staining. Here, we present a pipeline integrating three imaging modalities to visualize the same specimen at cryogenic temperature at different scales: cryo-fluorescence confocal microscopy, volume cryo-focused ion beam scanning electron microscopy, and transmission cryo-electron tomography. Our proof-of-concept benchmark revealed the 3D distribution of organelles and subcellular structures in whole heat-shocked yeast cells, including the ultrastructure of protein inclusions that recruit fluorescently-labeled chaperone Hsp104. Since our workflow efficiently integrates imaging at three different scales and can be applied to other types of cells, it could …
Mechanisms And Functions Of Spatial Protein Quality Control, Emily M. Sontag, Rahul S. Samant, Judith Frydman
Mechanisms And Functions Of Spatial Protein Quality Control, Emily M. Sontag, Rahul S. Samant, Judith Frydman
Biological Sciences Faculty Research and Publications
A healthy proteome is essential for cell survival. Protein misfolding is linked to a rapidly expanding list of human diseases, ranging from neurodegenerative diseases to aging and cancer. Many of these diseases are characterized by the accumulation of misfolded proteins in intra- and extracellular inclusions, such as amyloid plaques. The clear link between protein misfolding and disease highlights the need to better understand the elaborate machinery that manages proteome homeostasis, or proteostasis, in the cell. Proteostasis depends on a network of molecular chaperones and clearance pathways involved in the recognition, refolding, and/or clearance of aberrant proteins. Recent studies reveal that …
Identifying Polyglutamine Protein Species In Situ That Best Predict Neurodegeneration, Jason Miller, Montserrat Arrasate, Elizabeth Brooks, Clare Peter Libeu, Justin Legleiter, Danny Hatters, Jessica Curtis, Kenneth Cheung, Preethi Krishnan, Siddhartha Mitra, Kartika Widjaja, Benjamin A. Shaby, Gregor P. Lotz, Yvonne Newhouse, Emily M. Sontag, Alex Osmand, Michelle Gray, Vanitha Thulasiramin, Frederic Saudou, Mark Segal, X William Yang, Eliezer Masliah, Leslie M. Thompson, Paul J. Muchowski, Karl H. Weisgraber, Steven Finkbeiner
Identifying Polyglutamine Protein Species In Situ That Best Predict Neurodegeneration, Jason Miller, Montserrat Arrasate, Elizabeth Brooks, Clare Peter Libeu, Justin Legleiter, Danny Hatters, Jessica Curtis, Kenneth Cheung, Preethi Krishnan, Siddhartha Mitra, Kartika Widjaja, Benjamin A. Shaby, Gregor P. Lotz, Yvonne Newhouse, Emily M. Sontag, Alex Osmand, Michelle Gray, Vanitha Thulasiramin, Frederic Saudou, Mark Segal, X William Yang, Eliezer Masliah, Leslie M. Thompson, Paul J. Muchowski, Karl H. Weisgraber, Steven Finkbeiner
Biological Sciences Faculty Research and Publications
Polyglutamine (polyQ) stretches exceeding a threshold length confer a toxic function to proteins that contain them and cause at least nine neurological disorders. The basis for this toxicity threshold is unclear. Although polyQ expansions render proteins prone to aggregate into inclusion bodies, this may be a neuronal coping response to more toxic forms of polyQ. The exact structure of these more toxic forms is unknown. Here we show that the monoclonal antibody 3B5H10 recognizes a species of polyQ protein in situ that strongly predicts neuronal death. The epitope selectively appears among some of the many low-molecular-weight conformational states assumed by …
Fc177, A Minor Dec-1 Proprotein, Is Necessary To Prevent Ectopic Aggregation Of The Endochorion During Eggshell Assembly In Drosophila, Debra Kay Mauzy-Melitz, Gail L. Waring
Fc177, A Minor Dec-1 Proprotein, Is Necessary To Prevent Ectopic Aggregation Of The Endochorion During Eggshell Assembly In Drosophila, Debra Kay Mauzy-Melitz, Gail L. Waring
Biological Sciences Faculty Research and Publications
The Drosophila eggshell is a highly specialized extracellular matrix that forms between the oocyte and the surrounding epithelial follicle cells during late oogenesis. The dec-1 gene, which is required for proper eggshell assembly, produces three proproteins that are cleaved within the vitelline membrane layer to multiple derivatives. The different spatial distributions of the cleaved derivatives suggest that they play distinct roles in eggshell assembly. Using extant dec-1 mutations in conjunction with genetically engineered dec-1 transgenes, we show that, although all three dec-1 proproteins, fc106, fc125, and fc177, are required for female fertility, gross morphological abnormalities in the eggshell are observed …