A5040331827- Microtubule and mitosis dynamics
- Ubiquitin and proteasome pathways
- Genomics and Chromatin Dynamics
- RNA Interference and Gene Delivery
- Insect and Arachnid Ecology and Behavior
- Insect and Pesticide Research
- Cellular transport and secretion
- RNA Research and Splicing
- Mitochondrial Function and Pathology
- CAR-T cell therapy research
- DNA Repair Mechanisms
- CRISPR and Genetic Engineering
- Acute Myeloid Leukemia Research
- RNA and protein synthesis mechanisms
- Photosynthetic Processes and Mechanisms
- interferon and immune responses
- Cancer-related Molecular Pathways
- Advanced biosensing and bioanalysis techniques
- Monoclonal and Polyclonal Antibodies Research
- Cancer Genomics and Diagnostics
- Protein Degradation and Inhibitors
- DNA and Nucleic Acid Chemistry
- Protist diversity and phylogeny
- Plant nutrient uptake and metabolism
- Software Testing and Debugging Techniques
Fred Hutch Cancer Center
2012-2024
Cancer Research Center
2012-2022
Howard Hughes Medical Institute
2020-2022
Colorado State University
2012-2022
University of North Carolina at Greensboro
2020-2021
Hutchinson (United Kingdom)
2020
Western Washington University
2015
The Evergreen State College
2012
University College London
2012
Samsung Medical Center
2012
Abstract The existence of a 30‐nm fiber as basic folding unit for DNA packaging has remained topic active discussion. Here, we characterize the supramolecular structures formed by reversible Mg 2+ ‐dependent self‐association linear 12‐mer nucleosomal arrays using microscopy and physicochemical approaches. These reconstituted chromatin structures, which call “oligomers”, are globular throughout all stages cooperative assembly range in size from ~50 nm to maximum diameter ~1,000 nm. were...
To identify key regulators of human brain tumor maintenance and initiation, we performed multiple genome-wide RNAi screens in patient-derived glioblastoma multiforme (GBM) stem cells (GSCs). These identified the plant homeodomain (PHD)-finger domain protein PHF5A as differentially required for GSC expansion, compared with untransformed neural (NSCs) fibroblasts. Given PHF5A's known involvement facilitating interactions between U2 snRNP complex ATP-dependent helicases, examined...
Abstract To identify new candidate therapeutic targets for glioblastoma multiforme, we combined functional genetics and network modeling to kinases required the growth of patient-derived brain tumor–initiating cells (BTIC) but that are dispensable proliferating human neural stem (NSC). This approach yielded BUB1B/BUBR1, a critical mitotic spindle checkpoint player, as top-scoring lethal kinase. Knockdown BUB1B inhibited expansion BTIC isolates, both in vitro vivo, without affecting...
The H1 linker histones are abundant chromatin-associated DNA-binding proteins. Recent evidence suggests that also may function through protein–protein interactions. To gain a better understanding of the scope histone involvement in interactions, we used proteomics approach to identify H1-binding proteins human nuclear extracts. Full-length H1.0 and lacking its C-terminal domain (CTD) were for protein pull-downs. A total 107 candidate binding identified by LC-MS/MS. About one-third...
Highlights•BuGZ is a kinetochore protein that binds to and stabilizes Bub3•BuGZ localizes the Bub3 through conserved GLEBS domain•BuGZ depletion in transformed cells results severe chromosome alignment defects•Inhibiting Bub3's domain interactions may be therapeutic strategy for GBMSummaryDuring mitosis, spindle assembly checkpoint (SAC) monitors attachment of kinetochores (KTs) plus ends microtubules (MTs) prevents anaphase onset until chromosomes are aligned KTs under proper tension. Here,...
Accurate chromosome segregation requires kinetochores on duplicated chromatids to biorient by attaching dynamic microtubules from opposite spindle poles, which exerts forces bring under tension. However, initially bind indiscriminately, resulting in errors that must be corrected. While the Aurora B protein kinase destabilizes low-tension attachments phosphorylating kinetochores, are intrinsically less stable than those higher tension vitro independent of activity. Intrinsic tension-sensitive...
Glioblastoma multiforme (GBM) remains a mainly incurable disease in desperate need of more effective treatments. In this study, we develop evidence that the mitotic spindle checkpoint molecule BUB1B may offer predictive marker for aggressiveness and drug response. A subset GBM tumor isolates requires to suppress lethal kinetochore-microtubule attachment defects. Using gene expression data from stem-like cells, astrocytes, neural progenitor cells are sensitive or resistant inhibition, created...
In contrast to the western honey bee, Apis mellifera, other bee species have been largely neglected despite their importance and diversity. The genetic basis of evolutionary diversification bees remains unknown. Here, we provide a genome-wide comparison three species, each representing one subgenera bees, namely dwarf (Apis florea), giant (A. dorsata), cavity-nesting mellifera) with bumblebees as an outgroup. Our analyses resolve phylogeny diverging first. We find that evolution increased...
Aneuploidy, the incorrect number of whole chromosomes, is a common feature tumors that contributes to their initiation and evolution. Preventing aneuploidy requires properly functioning kinetochores, which are large protein complexes assembled on centromeric DNA link mitotic chromosomes dynamic spindle microtubules facilitate chromosome segregation. The kinetochore leverages at least two mechanisms prevent aneuploidy: error correction assembly checkpoint (SAC). BubR1, factor involved in both...
Kinetochores are large protein structures assembled on centromeric DNA during mitosis that bind to microtubules of the mitotic spindle orchestrate and power chromosome movements. Deregulation kinetochore-microtubule (KT-MT) attachments has been implicated in driving instability cancer evolution; however, nature source KT-MT attachment defects cells remain largely unknown. Here, we highlight recent findings suggesting oncogene-driven changes kinetochore regulation occur glioblastoma...
Zinc finger domain genes comprise ~3% of the human genome, yet many their functions remain unknown. Here we investigated roles for vertebrate-specific BTB zinc gene ZNF131 in context brain tumors. We report that is broadly required Glioblastoma stem-like cell (GSC) viability, but dispensable neural progenitor (NPC) viability. Examination expression changes after knockdown (kd) revealed activity notably promotes Joubert Syndrome ciliopathy genes, including KIF7, NPHP1, and TMEM237, as well...
Honey bees (Apis mellifera L) suffer from many brood pathogens, including viruses. Despite considerable research, the molecular responses and dynamics of honey bee pupae to viral pathogens remain poorly understood. Israeli Acute Paralysis Virus (IAPV) is emerging as a model virus since its association with severe colony losses. Using worker pupae, we studied transcriptomic methylomic consequences IAPV infection over three distinct time points after inoculation. Contrasts gene expression 5mC...
The factor VIII C2 domain is essential for binding to activated platelet surfaces as well the cofactor activity of in blood coagulation. Inhibitory antibodies against commonly develop following replacement therapy hemophilia A patients, or they may spontaneously arise cases acquired hemophilia. Porcine an effective therapeutic patients with inhibitor due its low cross-reactivity; however, molecular basis this behavior poorly understood. In study, X-ray crystal structure porcine was...
Aneuploidy, a condition that results from unequal partitioning of chromosomes during mitosis, is hallmark many cancers, including those caused by human papillomaviruses (HPVs). E6 and E7 are the primary transforming proteins in HPV drive tumor progression. In this study, we stably expressed noncancerous RPE1 cells analyzed specific mitotic defects contribute to aneuploidy each cell line. We find expression multiple associated with one or both spindle poles, causing significant delay. most...
Glioblastoma (GBM) is the most common and aggressive brain tumor in adults. To identify genes differentially required for viability of GBM stem-like cells (GSCs), we performed functional genomic lethality screens comparing GSCs control human neural stem cells. Among top-scoring hits a subset was F-box-containing gene FBXO42, which also predicted to be essential ∼15% cell lines derived from broad range cancers. Mechanistic studies revealed that, sensitive cells, FBXO42 activity prevents...
BuGZ is a kinetochore component that binds to and stabilizes Bub3, key player in mitotic spindle assembly checkpoint signaling. Bub3 required for recruitment of Bub1 BubR1, two proteins have essential distinct roles the checkpoint. Both BubR1 localize kinetochores through interactions with which are mediated conserved GLEBS domains both BubR1. also has domain, its localization as well, presumably binding. Although much understood about requirements interaction kinetochores, less known...
Abstract Background Adult and pediatric tumors display stark differences in their mutation spectra chromosome alterations. Here, we attempted to identify common unique gene dependencies associated biomarkers among adult tumor isolates using functional genetic lethal screens computational modeling. Methods We performed CRISRP-Cas9 lethality two glioblastoma (GBM) five brain representing atypical teratoid rhabdoid (ATRT), diffuse intrinsic pontine glioma, GBM, medulloblastoma. then integrated...
The identity of human protein-coding genes is well known, yet our in-depth knowledge their molecular functions and domain architecture remains limited by shortcomings in homology-based predictions experimental approaches focused on whole-gene depletion. To bridge this gap, we developed a method that leverages CRISPR-Cas9-induced mutations across for the priori identification functional regions at sequence level. As test case, applied to 48 mitotic genes, revealing hundreds required cell...
SUMMARY Kinetochores are large protein complexes that assemble at the centromere and bind to mitotic spindle microtubules ensure accurate chromosome segregation. Like most protein-coding genes, full multifunctional nature of kinetochore factors remains uncharacterized due limited experimental tools for unbiased dissection human sequences. We developed a method leverages CRISPR-Cas9 induced mutations identify key functional regions within sequences required cellular outgrowth. Our analysis 48...
Abstract Glioblastoma multiforme (GBM) is an aggressive and refractory cancer with limited therapeutic strategies poor survival. One challenge in treating this disease the ability of single glioblastoma stem-like cells (GSCs) to initiate tumors. To expand repertoire for GBM we performed RNAi screening GSCs a proposed cell origin, neural crest stem (NSCs). In these screens identified multiple regulators kinetochore-microtubule attachments as specifically required GSC Kinetochores are composed...