While the subcellular organisation of mitochondria is likely to influence many aspects cell physiology, its molecular control poorly understood. Here, we have investigated role retrograde motor protein complex, dynein-dynactin, in mitochondrial localisation and morphology. Disruption dynein function, achieved HeLa cells either by over-expressing dynactin subunit, dynamitin (p50), or microinjection an anti-dynein intermediate chain antibody, resulted (a) redistribution nuclear periphery, (b)...

A monoclonal antibody (M3A5), raised against microtubule-associated protein 2 (MAP-2), recognized an antigen associated with the Golgi complex in a variety of non-neuronal tissue culture cells. In double immunofluorescence studies M3A5 staining was very similar to that specific markers, even after disruption apparatus organization monensin or nocodazole. one band Mr approximately 110,000 immunoblots cell extracts; this protein, designated 110K, enriched stack fractions prepared from rat...

The mammalian Golgi complex is comprised of a ribbon stacked cisternal membranes often located in the pericentriolar region cell. Here, we report that during apoptosis fragmented into dispersed clusters tubulo-vesicular membranes. We have found fragmentation caspase dependent and identified GRASP65 (Golgi reassembly stacking protein 65 kD) as novel substrate. cleaved specifically by caspase-3 at conserved sites its membrane distal COOH terminus an early stage execution phase. Expression...

After viral fusion, capsids of the neurotropic herpes simplex virus are transported along microtubules (MT) to nuclear pores for genome uncoating, transcription and replication. assembly egress from nucleus, cytosolic host membranes secondary envelopment or axon terminal further spread. Using GFP‐tagged capsids, Cy3‐labelled MT cytosol, we have reconstituted capsid transport in vitro . In presence ATP, moved up 30 µm. Blocking function dynactin, a cofactor dynein kinesin‐2, inhibited...

Generating the extended endoplasmic reticulum (ER) network depends on microtubules, which act as tracks for motor-driven ER tubule movement, generate force to extend tubules by means of attachment growing microtubule plus-ends and provide static points. We have analysed dynamics in living VERO cells find that most extension is driven motors. Surprisingly, we observe ∼50% rapid movements occur direction centre cell, cytoplasmic dynein. Inhibition this movement leads an accumulation lamellar...

Cytoplasmic dynein is a minus end-directed microtubule motor that performs distinct functions in interphase and mitosis. In interphase, transports organelles along microtubules, whereas metaphase this has been implicated mitotic spindle formation orientation as well chromosome segregation. The manner which activity regulated during the cell cycle, however, not resolved. study, we have examined mechanism by organelle transport controlled cycle extracts of Xenopus laevis eggs. Here, show...

Recruitment of secretory vesicles to the cell surface is essential for sustained secretion insulin in response glucose. At present, molecular motors involved this movement, and mechanisms whereby they may be regulated, are undefined. To investigate role kinesin family members, we labelled densecore clonal β-cells using an adenovirally expressed, vesicle-targeted green fluorescent protein(phogrin.EGFP), employed immunoadsorption obtain highly purified insulin-containing vesicles. Whereas...

When higher eukaryotic cells enter mitosis, membrane organization changes dramatically and traffic between compartments is inhibited. Since transport along microtubules involved in secretion, endocytosis, the positioning of organelles during interphase, we have explored whether mitotic reorganization could involve a change microtubule-based transport. This question was examined by reconstituting organelle Xenopus egg extracts, which can be converted interphase metaphase states vitro absence...

Plasma membrane blebbing is a defining characteristic of apoptosis, but its significance not understood. Using live-cell imaging we have identified two phases apoptotic blebbing. The early phase restricted to adherent cells, and prevented by the Rho-activated kinase inhibitor Y27632. late partially resistant Y27632, generates morphologically distinct protrusions that are likely precursors bodies. Late observed in all cells tested. It occurs at fixed period before phosphatidyl serine...

LIS1, NDE1 and NDEL1 modulate cytoplasmic dynein function in several cellular contexts. However, evidence that they regulate dynein-dependent organelle positioning is limited. Here, we show depletion of or alone profoundly affected the organisation Golgi complex but did not cause it to disperse, slightly position endocytic compartments. striking dispersal organelles was observed when both were depleted. A substantial portion membrane associated, these proteins led complete loss from...

Summary— Detrusor instability has remained resistant to conventional forms of treatment. An attempt use biofeedback methods in its management is described. Six female patients with symptoms frequency, urgency and urge incontinence due detrusor were conditioned auditory visual stimuli for 6 81 h sessions. They assessed clinically urodynamically. The results are presented as well detailed case studies 3 patients. Subjectively, cured, 2 improved 1 the same; objectively, same. No significant...

Intracellular transport is predominantly heterogeneous in both time and space, exhibiting varying non-Brownian behavior. Characterization of this movement through averaging methods over an ensemble trajectories or the course a single trajectory often fails to capture heterogeneity. Here, we developed deep learning feedforward neural network trained on fractional Brownian motion, providing novel, accurate efficient method for resolving behavior intracellular space time. The requires...

Interphase Xenopus egg extracts form extensive tubular membrane networks in vitro. These are identified here as endoplasmic reticulum by the presence of ER resident proteins, shown immunofluorescence, and single ribosomes polysomes, electron microscopy. The effect phosphorylation on movement interphase was tested using phosphatase inhibitor, okadaic acid. Okadaic acid treatment resulted an increase up to 27-fold number tubules moving extent formed compared control extracts. This activation...

Cenp-F is an unusual kinetochore protein in that it localizes to the nuclear matrix interphase and envelope at G2/M transition; farnesylated rapidly degraded after mitosis. We have recently shown farnesylation of required for progression, its localization kinetochores, degradation. However, role plays mitosis has remained enigmatic. Here we show that, following repression by RNA interference (RNAi), processes metaphase chromosome alignment, anaphase segregation cytokinesis all fail. Although...