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https://www.readbyqxmd.com/read/28936467/peroxisome-motility-measurement-and-quantification-assay
#1
Jeremy Metz, Inês G Castro, Michael Schrader
Organelle movement, distribution and interaction contribute to the organisation of the eukaryotic cell. Peroxisomes are multifunctional organelles which contribute to cellular lipid metabolism and ROS homeostasis. They distribute uniformly in mammalian cells and move along microtubules via kinesin and dynein motors. Their metabolic cooperation with mitochondria and the endoplasmic reticulum (ER) is essential for the β-oxidation of fatty acids and the synthesis of myelin lipids and polyunsaturated fatty acids...
September 5, 2017: Bio-protocol
https://www.readbyqxmd.com/read/28933687/tagging-of-the-vaccinia-virus-protein-f13-with-mcherry-causes-aberrant-virion-morphogenesis
#2
David C J Carpentier, Michael S Hollinshead, Helen A Ewles, Stacey-Ann Lee, Geoffrey L Smith
Vaccinia virus produces two distinct infectious virions; the single-enveloped intracellular mature virus (IMV), which remains in the cell until cell lysis, and the double-enveloped extracellular enveloped virus (EEV), which mediates virus spread. The latter is derived from a triple-enveloped intracellular enveloped virus (IEV) precursor, which is transported to the cell periphery by the kinesin-1 motor complex. This transport involves the viral protein A36 as well as F12 and E2. A36 is an integral membrane protein associated with the outer virus envelope and is the only known direct link between virion and kinesin-1 complex...
September 20, 2017: Journal of General Virology
https://www.readbyqxmd.com/read/28930676/localized-phosphorylation-of-a-kinesin-1-adaptor-by-a-capsid-associated-kinase-regulates-hiv-1-motility-and-uncoating
#3
Viacheslav Malikov, Mojgan H Naghavi
Although microtubule motors mediate intracellular virus transport, the underlying interactions and control mechanisms remain poorly defined. This is particularly true for HIV-1 cores, which undergo complex, interconnected processes of cytosolic transport, reverse transcription, and uncoating of the capsid shell. Although kinesins have been implicated in regulating these events, curiously, there are no direct kinesin-core interactions. We recently showed that the capsid-associated kinesin-1 adaptor protein, fasciculation and elongation protein zeta-1 (FEZ1), regulates HIV-1 trafficking...
September 19, 2017: Cell Reports
https://www.readbyqxmd.com/read/28925357/restraint-of-presynaptic-protein-levels-by-wnd-dlk-signaling-mediates-synaptic-defects-associated-with-the-kinesin-3-motor-unc-104
#4
Jiaxing Li, Yao V Zhang, Elham Asghari Adib, Doychin T Stanchev, Xin Xiong, Susan Klinedinst, Pushpanjali Soppina, Thomas Robert Jahn, Richard I Hume, Tobias M Rasse, Catherine A Collins
The kinesin-3 family member Unc-104/KIF1A is required for axonal transport of many presynaptic components to synapses, and mutation of this gene results in synaptic dysfunction in mice, flies and worms. Our studies at the Drosophila neuromuscular junction indicate that many synaptic defects in unc-104-null mutants are mediated independently of Unc-104's transport function, via the Wallenda (Wnd)/DLK MAP kinase axonal damage signaling pathway. Wnd signaling becomes activated when Unc-104's function is disrupted, and leads to impairment of synaptic structure and function by restraining the expression level of active zone (AZ) and synaptic vesicle (SV) components...
September 19, 2017: ELife
https://www.readbyqxmd.com/read/28912692/calsyntenins-are-expressed-in-a-dynamic-and-partially-overlapping-manner-during-neural-development
#5
Gemma de Ramon Francàs, Tobias Alther, Esther T Stoeckli
Calsyntenins form a family of linker proteins between distinct populations of vesicles and kinesin motors for axonal transport. They were implicated in synapse formation and synaptic plasticity by findings in worms, mice and humans. These findings were in accordance with the postsynaptic localization of the Calsyntenins in the adult brain. However, they also affect the formation of neural circuits, as loss of Calsyntenin-1 (Clstn1) was shown to interfere with axonal branching and axon guidance. Despite the fact that Calsyntenins were discovered originally in embryonic chicken motoneurons, their distribution in the developing nervous system has not been analyzed in detail so far...
2017: Frontiers in Neuroanatomy
https://www.readbyqxmd.com/read/28901373/kinesin-and-dynein-mechanics-measurement-methods-and-research-applications
#6
Zachary Abraham, Emma Hawley, Daniel Hayosh, Victoria Webster-Wood, Ozan Akkus
Motor proteins play critical roles in the normal function of cells and proper development of organisms. Among motor proteins, failings in the normal function of two types of proteins, kinesin and dynein, have been shown to lead many pathologies, including neurodegenerative diseases and cancers. As such, it is critical for researchers to understand the underlying mechanics and behaviors of these proteins, not only to shed light on how failures may lead to disease, but also to guide research towards novel treatment and nanoengineering solutions...
September 13, 2017: Journal of Biomechanical Engineering
https://www.readbyqxmd.com/read/28892560/mutations-of-kif14-cause-primary-microcephaly-by-impairing-cytokinesis
#7
Abubakar Moawia, Ranad Shaheen, Sajida Rasool, Syeda Seema Waseem, Nour Ewida, Birgit Budde, Amit Kawalia, Susanne Motameny, Kamal Khan, Ambrin Fatima, Muhammad Jameel, Farid Ullah, Talia Akram, Zafar Ali, Uzma Abdullah, Saba Irshad, Wolfgang Höhne, Angelika Anna Noegel, Mohammed Al-Owain, Konstanze Hörtnagel, Petra Stöbe, Shahid Mahmood Baig, Peter Nürnberg, Fowzan Sami Alkuraya, Andreas Hahn, Muhammad Sajid Hussain
OBJECTIVE: Autosomal recessive primary microcephaly (MCPH) is a rare condition characterized by a reduced cerebral cortex accompanied with intellectual disability. Mutations in 17 genes have been shown to cause this phenotype. Recently, mutations in CIT, encoding CRIK (Citron Rho-interacting kinase) - a component of the central spindle matrix, were added. We aimed at identifying novel MCPH-associated genes and exploring their functional role in pathogenesis. METHODS: Linkage analysis and whole-exome sequencing were performed in consanguineous and nonconsanguineous MCPH families to identify disease-causing variants...
September 11, 2017: Annals of Neurology
https://www.readbyqxmd.com/read/28891968/biogenetic-relationships-of-bioactive-sponge-merotriterpenoids
#8
REVIEW
Thomas E Smith
Hydroquinone meroterpenoids, especially those derived from marine sponges, display a wide range of biological activities. However, use of these compounds is limited by their inaccessibility; there is no sustainable supply of these compounds. Furthermore, our knowledge of their metabolic origin remains completely unstudied. In this review, an in depth structural analysis of sponge merotriterpenoids, including the adociasulfate family of kinesin motor protein inhibitors, provides insight into their biosynthesis...
September 10, 2017: Marine Drugs
https://www.readbyqxmd.com/read/28883092/human-microcephaly-aspm-protein-is-a-spindle-pole-focusing-factor-that-functions-redundantly-with-cdk5rap2
#9
Elsa A Tungadi, Ami Ito, Tomomi Kiyomitsu, Gohta Goshima
Nonsense mutations in the ASPM gene have been most frequently identified among familial microcephaly patients. Depletion of the Drosophila orthologue causes spindle pole unfocusing during mitosis in multiple cell types. However, it remains unknown whether human ASPM has a similar function. Here, using CRISPR-based gene knockout (KO) and RNA interference combined with auxin-inducible degron, we show that ASPM functions in spindle pole organisation during mitotic metaphase redundantly with another microcephaly protein CDK5RAP2 (also called CEP215) in human tissue culture cells...
September 7, 2017: Journal of Cell Science
https://www.readbyqxmd.com/read/28860275/14-3-3-regulation-of-ncd-reveals-a-new-mechanism-for-targeting-proteins-to-the-spindle-in-oocytes
#10
Robin Beaven, Ricardo Nunes Bastos, Christos Spanos, Pierre Romé, C Fiona Cullen, Juri Rappsilber, Régis Giet, Gohta Goshima, Hiroyuki Ohkura
The meiotic spindle is formed without centrosomes in a large volume of oocytes. Local activation of crucial spindle proteins around chromosomes is important for formation and maintenance of a bipolar spindle in oocytes. We found that phosphodocking 14-3-3 proteins stabilize spindle bipolarity in Drosophila melanogaster oocytes. A critical 14-3-3 target is the minus end-directed motor Ncd (human HSET; kinesin-14), which has well-documented roles in stabilizing a bipolar spindle in oocytes. Phospho docking by 14-3-3 inhibits the microtubule binding activity of the nonmotor Ncd tail...
August 31, 2017: Journal of Cell Biology
https://www.readbyqxmd.com/read/28830772/covalent-and-non-covalent-chemical-engineering-of-actin-for-biotechnological-applications
#11
REVIEW
Saroj Kumar, Alf Mansson
The cytoskeletal filaments are self-assembled protein polymers with 8-25nm diameters and up to several tens of micrometres length. They have a range of pivotal roles in eukaryotic cells, including transportation of intracellular cargoes (primarily microtubules with dynein and kinesin motors) and cell motility (primarily actin and myosin) where muscle contraction is one example. For two decades, the cytoskeletal filaments and their associated motor systems have been explored for nanotechnological applications including miniaturized sensor systems and lab-on-a-chip devices...
August 19, 2017: Biotechnology Advances
https://www.readbyqxmd.com/read/28819981/highly-efficient-guiding-of-motile-microtubules-on-non-topographical-motor-patterns
#12
Cordula Reuther, Matthäus Mittasch, Sundar R Naganathan, Stephan W Grill, Stefan Diez
Molecular motors, highly efficient biological nanomachines, hold the potential to be employed for a wide range of nanotechnological applications. Toward this end, kinesin, dynein, or myosin motor proteins are commonly surface-immobilized within engineered environments in order to transport cargo attached to cytoskeletal filaments. Being able to flexibly control the direction of filament motion, and in particular on planar, non-topographical surfaces, has, however, remained challenging. Here, we demonstrate the applicability of a UV-laser-based ablation technique to programmably generate highly localized patterns of functional kinesin-1 motors with different shapes and sizes on PLL-g-PEG-coated polystyrene surfaces...
September 13, 2017: Nano Letters
https://www.readbyqxmd.com/read/28808088/the-armadillo-protein-p0071-controls-kif3-motor-transport
#13
Alexander Becher, Tim Eiseler, Marc Porzner, Paul Walther, René Keil, Susanne Bobrovich, Mechthild Hatzfeld, Thomas Seufferlein
We here report a novel function of the armadillo protein p0071 during KIF3/kinesin-2-mediated transport. Secretion of chromogranin A and matrix metallopeptidase 9 from pancreatic neuroendocrine tumor cells or pancreatic cancer cells, respectively, was substantially reduced following knockdown of p0071. Vesicle tracking indicated impaired directional persistence of vesicles upon p0071 depletion. This suggests a disturbed balance between plus- and minus-end directed microtubule transport in cells lacking p0071...
August 14, 2017: Journal of Cell Science
https://www.readbyqxmd.com/read/28805800/optogenetic-control-of-kinetochore-function
#14
Huaiying Zhang, Chanat Aonbangkhen, Ekaterina V Tarasovetc, Edward R Ballister, David M Chenoweth, Michael A Lampson
Kinetochores act as hubs for multiple activities during cell division, including microtubule interactions and spindle checkpoint signaling. Each kinetochore can act autonomously, and activities change rapidly as proteins are recruited to, or removed from, kinetochores. Understanding this dynamic system requires tools that can manipulate kinetochores on biologically relevant temporal and spatial scales. Optogenetic approaches have the potential to provide temporal and spatial control with molecular specificity...
October 2017: Nature Chemical Biology
https://www.readbyqxmd.com/read/28754069/axonal-spheroids-in-ovine-neuroaxonal-dystrophy-are-immunopositive-to-kinesin-and-dynein
#15
John W Finnie, Jim Manavis
Neuroaxonal dystrophy (NAD) is a neurologic disorder of sheep characterized by accumulation of numerous axonal swellings (spheroids) in specific regions of the brainstem and spinal cord. Disruption of axonal transport, which is driven in anterograde and retrograde directions by the molecular motors, kinesin and dynein, respectively, is believed to contribute to spheroid development. Accordingly, we examined spheroids in ovine NAD cases immunohistochemically for kinesin and dynein and found both motor proteins, with dynein more strongly expressed than kinesin...
July 1, 2017: Journal of Veterinary Diagnostic Investigation
https://www.readbyqxmd.com/read/28747564/high-eg5-expression-predicts-poor-prognosis-in-breast-cancer
#16
Qin Jin, Fang Huang, Xudong Wang, Huijun Zhu, Yun Xian, Jieying Li, Shu Zhang, Qichao Ni
Eg5 is a motor protein belonging to the kinesin-5 family and has been suggested to exert important function in tumors. In this study, we determined the mRNA and protein expression levels of Eg5 in cancerous and non-cancerous breast tissue by quantitative real-time polymerase chain reaction (qRT-PCR) and tissue microarray immunohistochemistry analysis (TMA-IHC) respectively. The results of 20 fresh-frozen BC samples demonstrated that Eg5 mRNA levels were significantly higher in BC tissues compared with corresponding non-cancerous tissue (p = 0...
July 10, 2017: Oncotarget
https://www.readbyqxmd.com/read/28736169/protein-interaction-analysis-provides-a-map-of-the-spatial-and-temporal-organization-of-the-ciliary-gating-zone
#17
Daisuke Takao, Liang Wang, Allison Boss, Kristen J Verhey
The motility and signaling functions of the primary cilium require a unique protein and lipid composition that is determined by gating mechanisms localized at the base of the cilium. Several protein complexes localize to the gating zone and may regulate ciliary protein composition; however, the mechanisms of ciliary gating and the dynamics of the gating components are largely unknown. Here, we used the BiFC (bimolecular fluorescence complementation) assay and report for the first time on the protein-protein interactions that occur between ciliary gating components and transiting cargoes during ciliary entry...
August 7, 2017: Current Biology: CB
https://www.readbyqxmd.com/read/28733669/high-resolution-imaging-of-a-single-gliding-protofilament-of-tubulins-by-hs-afm
#18
Jakia Jannat Keya, Daisuke Inoue, Yuki Suzuki, Toshiya Kozai, Daiki Ishikuro, Noriyuki Kodera, Takayuki Uchihashi, Arif Md Rashedul Kabir, Masayuki Endo, Kazuki Sada, Akira Kakugo
In vitro gliding assay of microtubules (MTs) on kinesins has provided us with valuable biophysical and chemo-mechanical insights of this biomolecular motor system. Visualization of MTs in an in vitro gliding assay has been mainly dependent on optical microscopes, limited resolution of which often render them insufficient sources of desired information. In this work, using high speed atomic force microscopy (HS-AFM), which allows imaging with higher resolution, we monitored MTs and protofilaments (PFs) of tubulins while gliding on kinesins...
July 21, 2017: Scientific Reports
https://www.readbyqxmd.com/read/28731566/heterogeneity-in-kinesin-function
#19
Babu J N Reddy, Suvranta Tripathy, Michael Vershinin, Marvin E Tanenbaum, Jing Xu, Michelle Mattson-Hoss, Karim Arabi, Dail Chapman, Tory Doolin, Changbong Hyeon, Steven P Gross
The kinesin family proteins are often studied as prototypical molecular motors; a deeper understanding of them can illuminate regulation of intracellular transport. It is typically assumed that they function identically. Here we find that this assumption of homogeneous function appears incorrect: variation among motors' velocities in vivo and in vitro is larger than the stochastic variation expected for an ensemble of "identical" motors. When moving on microtubules, slow and fast motors are persistently slow, and fast, respectively...
July 21, 2017: Traffic
https://www.readbyqxmd.com/read/28720664/dynein-efficiently-navigates-the-dendritic-cytoskeleton-to-drive-the-retrograde-trafficking-of-bdnf-trkb-signaling-endosomes
#20
Swathi Ayloo, Pedro Guedes-Dias, Amy E Ghiretti, Erika L F Holzbaur
The efficient transport of cargoes within axons and dendrites is critical for neuronal function. Although we have a basic understanding of axonal transport, much less is known about transport in dendrites. We used an optogenetic approach to recruit motor proteins to cargo in real time within axons or dendrites in hippocampal neurons. Kinesin-1, a robust axonal motor, moves cargo less efficiently in dendrites. In contrast, cytoplasmic dynein efficiently navigates both axons and dendrites; in both compartments, dynamic microtubule plus ends enhance dynein-dependent transport...
September 15, 2017: Molecular Biology of the Cell
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