A5054054349- Microtubule and mitosis dynamics
- Genetic and Kidney Cyst Diseases
- Protist diversity and phylogeny
- Photosynthetic Processes and Mechanisms
- Fungal and yeast genetics research
- Microbial Community Ecology and Physiology
- Renal and related cancers
- Micro and Nano Robotics
- Methane Hydrates and Related Phenomena
- Prenatal Screening and Diagnostics
- RNA Research and Splicing
- Plant nutrient uptake and metabolism
- Plant Molecular Biology Research
- Genomics and Chromatin Dynamics
- Cell Image Analysis Techniques
- Chromosomal and Genetic Variations
- Ubiquitin and proteasome pathways
- Advanced Electron Microscopy Techniques and Applications
- Epigenetics and DNA Methylation
- Protein Structure and Dynamics
- Connective tissue disorders research
- Signaling Pathways in Disease
- ATP Synthase and ATPases Research
- Advanced Fluorescence Microscopy Techniques
- Genomics and Phylogenetic Studies
University of Colorado Anschutz Medical Campus
2016-2025
University of Colorado Boulder
2006-2022
University of Colorado Denver
2009-2022
University of California, San Francisco
2022
University of Colorado Colorado Springs
2021
Pearson (United States)
2021
University of North Carolina at Chapel Hill
2001-2005
North Carolina State University
2003
The ability of kinetochores to recruit microtubules, generate force, and activate the mitotic spindle checkpoint may all depend on microtubule- and/or tension-dependent changes in kinetochore assembly. With use quantitative digital imaging immunofluorescence microscopy PtK1 tissue cells, we find that outer domain kinetochore, but not CREST-stained inner core, exhibits three microtubule-dependent assembly states, directly dependent tension. First, prometaphase with few or no microtubules have...
Using green fluorescent protein probes and rapid acquisition of high-resolution fluorescence images, sister centromeres in budding yeast are found to be separated oscillate between spindle poles before anaphase B elongation. The rates movement during these oscillations similar those microtubule plus end dynamics. degree preanaphase separation varies widely, with infrequent centromere reassociations observed anaphase. Centromeres a metaphase-like conformation, whereas chromosome arms neither...
Basal bodies organize the nine doublet microtubules found in cilia. Cilia are required for a variety of cellular functions, including motility and sensing stimuli. Understanding this biochemically complex organelle requires an inventory molecular components contribution each makes to overall structure. We define basal body proteome determine specific localization ciliated protozoan Tetrahymena thermophila. Using biochemical, bioinformatic, genetic approach, we identify 97 known candidate...
Centrioles are the foundation for centrosome and cilia formation. The biogenesis of centrioles is initiated by an assembly mechanism that first synthesizes ninefold symmetrical cartwheel subsequently leads to a stable cylindrical microtubule scaffold capable withstanding microtubule-based forces generated centrosomes cilia. We report conserved WD40 repeat domain–containing protein Poc1 required structural maintenance in Tetrahymena thermophila. Furthermore, human Poc1B primary ciliogenesis,...
Abstract Background Ichthyophthirius multifiliis , commonly known as Ich, is a highly pathogenic ciliate responsible for 'white spot', disease causing significant economic losses to the global aquaculture industry. Options control are extremely limited, and Ich's obligate parasitic lifestyle makes experimental studies challenging. Unlike most well-studied protozoan parasites, Ich belongs phylum composed primarily of free-living members. Indeed, it closely related model organism Tetrahymena...
Stu2p is a member of conserved family microtubule-binding proteins and an essential protein in yeast. Here, we report the first vivo analysis microtubule dynamics cells lacking this family. For these studies, have used conditional depletion strain expressing α-tubulin fused to green fluorescent protein. Depletion leads fewer less dynamic cytoplasmic microtubules both G1 preanaphase cells. The reduction due primarily decreases catastrophe rescue frequencies increase fraction time spend...
During metaphase in budding yeast mitosis, sister kinetochores are tethered to opposite poles and separated, stretching their intervening chromatin, by singly attached kinetochore microtubules (kMTs). Kinetochore movements coupled single microtubule plus-end polymerization/depolymerization at attachment sites. Here, we use computer modeling test possible mechanisms controlling chromosome alignment during simulating experiments that determine the 1) mean positions of Cse4-GFP, 2) extent...
Cilia, essential motile and sensory organelles, have several compartments: the basal body, transition zone, middle distal axoneme segments. The segment accommodates key functions, including cilium assembly activities. While contains doublet microtubules (incomplete B-tubules fused to complete A-tubules), only A-tubule extensions, its existence requires coordination of microtubule length at nanometer scale. We show that three conserved proteins, two which are mutated in ciliopathy Joubert...
Basal bodies (BBs) are conserved eukaryotic structures that organize cilia. They comprised of nine, cylindrically arranged, triplet microtubules (TMTs) connected to each other by inter-TMT linkages which stabilize the structure. Poc1 is a protein important for BB structural integrity in face ciliary forces transmitted BBs. To understand how confers stability, we identified precise position Tetrahymena and effect loss on binds at TMT inner junctions, stabilizing TMTs directly. From this...
It has been hypothesized that spatial gradients in kMT dynamic instability facilitate mitotic spindle formation and chromosome movement. To test this hypothesis requires the analysis of dynamics, which have not resolved at single level living cells. The budding yeast offers an attractive system to study dynamics because, contrast animal cells, there is only one per kinetochore. visualize metaphase plus-end yeast, a strain containing green fluorescent protein fusion kinetochore protein, Cse4,...
Basal bodies nucleate, anchor, and organize cilia. As the anchor for motile cilia, basal must be resistant to forces directed toward cell as a consequence of ciliary beating. The molecules generalized mechanisms that contribute maintenance remain discovered. Bld10/Cep135 is body outer cartwheel domain protein has established roles in assembly nascent bodies. We find Bld10 first incorporates stably at early during new assembly. continues accumulate after assembly, we hypothesize full...
Trisomy 21, the genetic cause of Down syndrome, disrupts primary cilia formation and function, in part through elevated Pericentrin, a centrosome protein encoded on chromosome 21. Yet how trisomy 21 Pericentrin disrupt cilia-related molecules pathways, vivo phenotypic relevance remain unclear. Utilizing ciliogenesis time course experiments combined with light microscopy electron tomography, we reveal that polyploidy elevates microtubules away from corral MyosinVA EHD1, delaying ciliary...
The interaction of kinetochores with dynamic microtubules during mitosis is essential for proper centromere motility, congression to the metaphase plate, and subsequent anaphase chromosome segregation. Budding yeast has been critical in discovery proteins necessary this interaction. However, molecular mechanism microtubule-kinetochore interactions remains poorly understood. Using live cell imaging mutations affecting microtubule binding kinetochore function, we identify a regulatory spindle...
Cilia-organizing basal bodies (BBs) are microtubule scaffolds that visibly asymmetrical because they have attached auxiliary structures, such as striated fibers. In multiciliated cells, BB orientation aligns to ensure coherent ciliary beating, but the mechanisms maintain unclear. For first time in Tetrahymena thermophila, we use comparative whole-genome sequencing identify mutation disorientation mutant disA-1. disA-1 abolishes localization of novel protein DisAp T. thermophila fibers...
Extracellular vesicles (EVs) are key contributors to cancer where they play an integral role in cell-cell communication and transfer pro-oncogenic molecules recipient cells thereby conferring a cancerous phenotype. Here, we purified EVs using straightforward biochemical approaches from multiple cell lines subsequently characterized these via biophysical methods. In addition, used fluorescence microscopy directly show internalization of into the within few minutes upon addition cells. We...
// Agnieszka A. Kendrick 1 , Johnathon Schafer Monika Dzieciatkowska Travis Nemkov Angelo D’Alessandro Deepika Neelakantan 2 Heide L. Ford Chad G. Pearson 3 Colin D. Weekes 4 Kirk C. Hansen Elan Z. Eisenmesser Department of Biochemistry and Molecular Genetics, School Medicine, University Colorado Denver, CO, USA Pharmacology, Cell Developmental Biology, Division Oncology, Correspondence to: Eisenmesser, email: elan.eisenmesser@ucdenver.edu Keywords: metabolism, ancillary protein, PDAC,...