Dynamics of myosin, microtubules, and Kinesin-6 at the cortex during cytokinesis in Drosophila S2 cells

© The Authors, 2009 . This article is distributed under the terms of the Creative Commons Attribution-Noncommercial-Share Alike 3.0 Unported License. The definitive version was published in Journal of Cell Biology 186 (2009): 727-738, doi:10.1083/jcb.200902083.Signals from the mitotic spindle during anaphase specify the location of the actomyosin contractile ring during cytokinesis, but the detailed mechanism remains unresolved. Here, we have imaged the dynamics of green fluorescent protein–tagged myosin filaments, microtubules, and Kinesin-6 (which carries activators of Rho guanosine triphosphatase) at the cell cortex using total internal reflection fluorescence microscopy in flattened Drosophila S2 cells. At anaphase onset, Kinesin-6 relocalizes to microtubule plus ends that grow toward the cortex, but refines its localization over time so that it concentrates on a subset of stable microtubules and along a diffuse cortical band at the equator. The pattern of Kinesin-6 localization closely resembles where new myosin filaments appear at the cortex by de novo assembly. While accumulating at the equator, myosin filaments disappear from the poles of the cell, a process that also requires Kinesin-6 as well as possibly other signals that emanate from the elongating spindle. These results suggest models for how Kinesin-6 might define the position of cortical myosin during cytokinesis.This work was supported by a National Institutes of Health grant NIH 38499 to R.D. Vale

Insights into Analogy Completion from the Biomedical Domain

Analogy completion has been a popular task in recent years for evaluating the semantic properties of word embeddings, but the standard methodology makes a number of assumptions about analogies that do not always hold, either in recent benchmark datasets or when expanding into other domains. Through an analysis of analogies in the biomedical domain, we identify three assumptions: that of a Single Answer for any given analogy, that the pairs involved describe the Same Relationship, and that each pair is Informative with respect to the other. We propose modifying the standard methodology to relax these assumptions by allowing for multiple correct answers, reporting MAP and MRR in addition to accuracy, and using multiple example pairs. We further present BMASS, a novel dataset for evaluating linguistic regularities in biomedical embeddings, and demonstrate that the relationships described in the dataset pose significant semantic challenges to current word embedding methods.Comment: Accepted to BioNLP 2017. (10 pages

Characterizing the impact of geometric properties of word embeddings on task performance

Analysis of word embedding properties to inform their use in downstream NLP tasks has largely been studied by assessing nearest neighbors. However, geometric properties of the continuous feature space contribute directly to the use of embedding features in downstream models, and are largely unexplored. We consider four properties of word embedding geometry, namely: position relative to the origin, distribution of features in the vector space, global pairwise distances, and local pairwise distances. We define a sequence of transformations to generate new embeddings that expose subsets of these properties to downstream models and evaluate change in task performance to understand the contribution of each property to NLP models. We transform publicly available pretrained embeddings from three popular toolkits (word2vec, GloVe, and FastText) and evaluate on a variety of intrinsic tasks, which model linguistic information in the vector space, and extrinsic tasks, which use vectors as input to machine learning models. We find that intrinsic evaluations are highly sensitive to absolute position, while extrinsic tasks rely primarily on local similarity. Our findings suggest that future embedding models and post-processing techniques should focus primarily on similarity to nearby points in vector space.Comment: Appearing in the Third Workshop on Evaluating Vector Space Representations for NLP (RepEval 2019). 7 pages + reference

USP45 and Spindly are part of the same complex implicated in cell migration

Abstract Ubiquitylation is a protein modification implicated in several cellular processes. This process is reversible by the action of deubiquinating enzymes (DUBs). USP45 is a ubiquitin specific protease about which little is known, aside from roles in DNA damage repair and differentiation of the vertebrate retina. Here, by using mass spectrometry we have identified Spindly as a new target of USP45. Our data show that Spindly and USP45 are part of the same complex and that their interaction specifically depends on the catalytic activity of USP45. In addition, we describe the type of ubiquitin chains associated with the complex that can be cleaved by USP45, with a preferential activity on K48 ubiquitin chain type and potentially K6. Here, we also show that Spindly is mono-ubiquitylated and this can be specifically removed by USP45 in its active form but not by the catalytic inactive form. Lastly, we identified a new role for USP45 in cell migration, similar to that which was recently described for Spindly

City of Columbus Green Memo III: Municipal Building Energy Management

Course Code: ENR/AEDE 4567Green Memo III proposed sustainability objectives for the City of Columbus, Ohio to reach by 2020. This project evaluates goals to reduce energy consumption in municipal facilities through ENERGY STAR energy monitoring software and the implementation of Energy Conservation Measures.Academic Major: Environment, Economy, Development, and Sustainabilit

Mechanoaccumulative elements of the mammalian actin cytoskeleton

To change shape, divide, form junctions, and migrate, cells reorganize their cytoskeletons in response to changing mechanical environments [1-4]. Actin cytoskeletal elements, including myosin II motors and actin crosslinkers, structurally remodel and activate signaling pathways in response to imposed stresses [5-9]. Recent studies demonstrate the importance of force-dependent structural rearrangement of α-catenin in adherens junctions [10] and vinculin's molecular clutch mechanism in focal adhesions [11]. However, the complete landscape of cytoskeletal mechanoresponsive proteins and the mechanisms by which these elements sense and respond to force remain to be elucidated. To find mechanosensitive elements in mammalian cells, we examined protein relocalization in response to controlled external stresses applied to individual cells. Here, we show that non-muscle myosin II, α-actinin, and filamin accumulate to mechanically stressed regions in cells from diverse lineages. Using reaction-diffusion models for force-sensitive binding, we successfully predicted which mammalian α-actinin and filamin paralogs would be mechanoaccumulative. Furthermore, a Goldilocks zone must exist for each protein where the actin-binding affinity must be optimal for accumulation. In addition, we leveraged genetic mutants to gain a molecular understanding of the mechanisms of α-actinin and filamin catch-bonding behavior. Two distinct modes of mechanoaccumulation can be observed: a fast, diffusion-based accumulation and a slower, myosin II-dependent cortical flow phase that acts on proteins with specific binding lifetimes. Finally, we uncovered cell-type and cell-cycle-stage-specific control of the mechanosensation of myosin IIB, but not myosin IIA or IIC. Overall, these mechanoaccumulative mechanisms drive the cell's response to physical perturbation during proper tissue development and disease

Requirement of the Dynein-Adaptor Spindly for Mitotic and Post-Mitotic Functions in Drosophila

Spindly was originally identified as a specific regulator of Dynein activity at the kinetochore. In early prometaphase, Spindly recruits the Dynein/Dynactin complex, promoting the establishment of stable kinetochore-microtubule interactions and progression into anaphase. While details of Spindly function in mitosis have been worked out in cultured human cells and in the C. elegans zygote, the function of Spindly within the context of an organism has not yet been addressed. Here, we present loss- and gain-of-function studies of Spindly using transgenic RNAi in Drosophila. Knock-down of Spindly in the female germ line results in mitotic arrest during embryonic cleavage divisions. We investigated the requirements of Spindly protein domains for its localisation and function, and found that the carboxy-terminal region controls Spindly localisation in a cell-type specific manner. Overexpression of Spindly in the female germ line is embryonic lethal and results in altered egg morphology. To determine whether Spindly plays a role in post-mitotic cells, we altered Spindly protein levels in migrating cells and found that ovarian border cell migration is sensitive to the levels of Spindly protein. Our study uncovers novel functions of Spindly and a differential, functional requirement for its carboxy-terminal region in Drosophila

Automated Coding of Under-Studied Medical Concept Domains: Linking Physical Activity Reports to the International Classification of Functioning, Disability, and Health

Linking clinical narratives to standardized vocabularies and coding systems is a key component of unlocking the information in medical text for analysis. However, many domains of medical concepts lack well-developed terminologies that can support effective coding of medical text. We present a framework for developing natural language processing (NLP) technologies for automated coding of under-studied types of medical information, and demonstrate its applicability via a case study on physical mobility function. Mobility is a component of many health measures, from post-acute care and surgical outcomes to chronic frailty and disability, and is coded in the International Classification of Functioning, Disability, and Health (ICF). However, mobility and other types of functional activity remain under-studied in medical informatics, and neither the ICF nor commonly-used medical terminologies capture functional status terminology in practice. We investigated two data-driven paradigms, classification and candidate selection, to link narrative observations of mobility to standardized ICF codes, using a dataset of clinical narratives from physical therapy encounters. Recent advances in language modeling and word embedding were used as features for established machine learning models and a novel deep learning approach, achieving a macro F-1 score of 84% on linking mobility activity reports to ICF codes. Both classification and candidate selection approaches present distinct strengths for automated coding in under-studied domains, and we highlight that the combination of (i) a small annotated data set; (ii) expert definitions of codes of interest; and (iii) a representative text corpus is sufficient to produce high-performing automated coding systems. This study has implications for the ongoing growth of NLP tools for a variety of specialized applications in clinical care and research.Comment: Updated final version, published in Frontiers in Digital Health, https://doi.org/10.3389/fdgth.2021.620828. 34 pages (23 text + 11 references); 9 figures, 2 table

Interview with James Figgs, civil rights activist and DJ

Interviewer and Recordist: Miles Laseter. Recorded at KBUD (Radio station : Sardis, Miss.