scholarly journals Binding of microtubule-associated protein 1B to LIS1 affects the interaction between dynein and LIS1

2005 ◽  
Vol 389 (2) ◽  
pp. 333-341 ◽  
Author(s):  
Eva M. JiméNez-Mateos ◽  
Francisco Wandosell ◽  
Orly Reiner ◽  
Jesús Avila ◽  
Christian González-Billault
Keyword(s):  
Neuronal Migration ◽  
Hippocampal Neurons ◽  
Morphological Changes ◽  
Molecular Motor ◽  
Cytoskeletal Proteins ◽  
Actin Binding ◽  
Microtubule Associated Proteins ◽  
Neural Processes ◽  
Associated Proteins ◽  
Differential Binding

For neuronal migration to occur, the cell must undergo morphological changes that require modifications of the cytoskeleton. Several different MAPs (microtubule-associated proteins) or actin-binding proteins are proposed to be involved in the migration of neurons. Therefore we have specifically analysed how two members of the MAP family, MAP1B and LIS1 (lissencephaly-related protein 1), interact with one another and participate in neuronal migration. Our results indicate that, in hippocampal neurons, MAP1B and LIS1 co-localize, associate and interact with each another. The interaction between these two MAPs is regulated by the phosphorylation of MAP1B. Furthermore, this interaction interferes with the association between LIS1 and the microtubule-dependent molecular motor, dynein. Clearly, the differential binding of these cytoskeletal proteins could regulate the functions attributed to the LIS1–dynein complex, including those related to extension of the neural processes necessary for neuronal migration.

scholarly journals Defects in Axonal Elongation and Neuronal Migration in Mice with Disrupted tau and map1b Genes

2000 ◽  
Vol 150 (5) ◽  
pp. 989-1000 ◽  
Author(s):  
Yosuke Takei ◽  
Junlin Teng ◽  
Akihiro Harada ◽  
Nobutaka Hirokawa
Keyword(s):  
Knockout Mice ◽  
Neuronal Migration ◽  
Hippocampal Neurons ◽  
Primary Cultures ◽  
Double Knockout ◽  
Microtubule Organization ◽  
Axonal Elongation ◽  
Microtubule Associated Proteins ◽  
Associated Proteins

Tau and MAP1B are the main members of neuronal microtubule-associated proteins (MAPs), the functions of which have remained obscure because of a putative functional redundancy (Harada, A., K. Oguchi, S. Okabe, J. Kuno, S. Terada, T. Ohshima, R. Sato-Yoshitake, Y. Takei, T. Noda, and N. Hirokawa. 1994. Nature. 369:488–491; Takei, Y., S. Kondo, A. Harada, S. Inomata, T. Noda, and N. Hirokawa. 1997. J. Cell Biol. 137:1615–1626). To unmask the role of these proteins, we generated double-knockout mice with disrupted tau and map1b genes and compared their phenotypes with those of single-knockout mice. In the analysis of mice with a genetic background of predominantly C57Bl/6J, a hypoplastic commissural axon tract and disorganized neuronal layering were observed in the brains of the tau+/+map1b−/− mice. These phenotypes are markedly more severe in tau−/−map1b−/− double mutants, indicating that tau and MAP1B act in a synergistic fashion. Primary cultures of hippocampal neurons from tau−/−map1b−/− mice showed inhibited axonal elongation. In these cells, a generation of new axons via bundling of microtubules at the neck of the growth cones appeared to be disturbed. Cultured cerebellar neurons from tau−/−map1b−/− mice showed delayed neuronal migration concomitant with suppressed neurite elongation. These findings indicate the cooperative functions of tau and MAP1B in vivo in axonal elongation and neuronal migration as regulators of microtubule organization.

scholarly journals NCAM2 Regulates Dendritic and Axonal Differentiation through the Cytoskeletal Proteins MAP2 and 14-3-3

2020 ◽  
Vol 30 (6) ◽  
pp. 3781-3799
Author(s):  
Antoni Parcerisas ◽  
Lluís Pujadas ◽  
Alba Ortega-Gascó ◽  
Bartomeu Perelló-Amorós ◽  
Ricardo Viais ◽  
...  
Keyword(s):  
Cell Biology ◽  
Hippocampal Neurons ◽  
Cytoskeletal Proteins ◽  
In Vivo Studies ◽  
Microtubule Associated Proteins ◽  
Neural Cell Adhesion ◽  
Aberrant Phenotype ◽  
Associated Proteins ◽  
And Migration

Abstract Neural cell adhesion molecule 2 (NCAM2) is involved in the development and plasticity of the olfactory system. Genetic data have implicated the NCAM2 gene in neurodevelopmental disorders including Down syndrome and autism, although its role in cortical development is unknown. Here, we show that while overexpression of NCAM2 in hippocampal neurons leads to minor alterations, its downregulation severely compromises dendritic architecture, leading to an aberrant phenotype including shorter dendritic trees, retraction of dendrites, and emergence of numerous somatic neurites. Further, our data reveal alterations in the axonal tree and deficits in neuronal polarization. In vivo studies confirm the phenotype and reveal an unexpected role for NCAM2 in cortical migration. Proteomic and cell biology experiments show that NCAM2 molecules exert their functions through a protein complex with the cytoskeletal-associated proteins MAP2 and 14-3-3γ and ζ. We provide evidence that NCAM2 depletion results in destabilization of the microtubular network and reduced MAP2 signal. Our results demonstrate a role for NCAM2 in dendritic formation and maintenance, and in neural polarization and migration, through interaction of NCAM2 with microtubule-associated proteins.

scholarly journals RhoA/ROCK regulation of neuritogenesis via profilin IIa–mediated control of actin stability

2003 ◽  
Vol 162 (7) ◽  
pp. 1267-1279 ◽  
Author(s):  
Jorge Santos Da Silva ◽  
Miguel Medina ◽  
Cecilia Zuliani ◽  
Alessia Di Nardo ◽  
Walter Witke ◽  
...  
Keyword(s):  
Neuronal Differentiation ◽  
Hippocampal Neurons ◽  
Morphological Changes ◽  
Decisive Role ◽  
Central Mechanism ◽  
Actin Binding ◽  
Actin Binding Protein ◽  
Rhoa Activity ◽  
Neuronal Soma ◽  
Cultured Hippocampal Neurons

Neuritogenesis, the first step of neuronal differentiation, takes place as nascent neurites bud from the immediate postmitotic neuronal soma. Little is known about the mechanisms underlying the dramatic morphological changes that characterize this event. Here, we show that RhoA activity plays a decisive role during neuritogenesis of cultured hippocampal neurons by recruiting and activating its specific kinase ROCK, which, in turn, complexes with profilin IIa. We establish that this previously uncharacterized brain-specific actin-binding protein controls neurite sprouting by modifying actin stability, a function regulated by ROCK-mediated phosphorylation. Furthermore, we determine that this novel cascade is switched on or off by physiological stimuli. We propose that RhoA/ROCK/PIIa-mediated regulation of actin stability, shown to be essential for neuritogenesis, may constitute a central mechanism throughout neuronal differentiation.

scholarly journals Intracellular Proadrenomedullin-Derived Peptides Decorate the Microtubules and Contribute to Cytoskeleton Function

Endocrinology ◽  
2008 ◽  
Vol 149 (6) ◽  
pp. 2888-2898 ◽  
Author(s):  
Dan L. Sackett ◽  
Laurent Ozbun ◽  
Enrique Zudaire ◽  
Lisa Wessner ◽  
John M. Chirgwin ◽  
...  
Keyword(s):  
Posttranslational Modifications ◽  
Morphological Changes ◽  
Microtubule Associated Proteins ◽  
Microtubule Stabilization ◽  
Gene Coding ◽  
Intracellular Compartments ◽  
Associated Proteins ◽  
G2 Phase ◽  
Interfering Rna

Adrenomedullin (AM) and proadrenomedullin N-terminal 20 peptide (PAMP) are secretory hormones, but it is not unusual to find them in intracellular compartments. Using yeast-2 hybrid technology, we found interactions between AM and several microtubule-associated proteins (MAPs), and between PAMP and tubulin. Expression of fluorescent-tagged AM and PAMP as well as immunofluorescence for the native peptides showed a complete decoration of the microtubules and colocalization with other MAPs. PAMP, but not AM, bound to tubulin in vitro and destabilized tubulin polymerization. Down-regulation of the gene coding for both AM and PAMP through small interfering RNA technology resulted in morphological changes, microtubule stabilization, increase in posttranslational modifications of tubulin such as acetylation and detyrosination, reduction in cell motility, and partial arrest at the G2 phase of the cell cycle, when compared with cells transfected with the same vector carrying a scrambled sequence. These results show that PAMP is a novel MAP, whereas AM may be exerting more subtle effects in regulating cytoskeleton function.

scholarly journals Cytoplasmic Dynein Mediates Adenovirus Binding to Microtubules

2004 ◽  
Vol 78 (18) ◽  
pp. 10122-10132 ◽  
Author(s):  
Samir A. Kelkar ◽  
K. Kevin Pfister ◽  
Ronald G. Crystal ◽  
Philip L. Leopold
Keyword(s):  
Molecular Motor ◽  
Retrograde Transport ◽  
Cytoplasmic Dynein ◽  
Bovine Brain ◽  
Microtubule Binding ◽  
Microtubule Associated Proteins ◽  
Primary Mode ◽  
Cell Lysate ◽  
Brain Maps ◽  
Associated Proteins

ABSTRACT During infection, adenovirus (Ad) capsids undergo microtubule-dependent retrograde transport as part of a program of vectorial transport of the viral genome to the nucleus. The microtubule-associated molecular motor, cytoplasmic dynein, has been implicated in the retrograde movement of Ad. We hypothesized that cytoplasmic dynein constituted the primary mode of association of Ad with microtubules. To evaluate this hypothesis, an Ad-microtubule binding assay was established in which microtubules were polymerized with taxol, combined with Ad in the presence or absence of microtubule-associated proteins (MAPs), and centrifuged through a glycerol cushion. The addition of purified bovine brain MAPs increased the fraction of Ad in the microtubule pellet from 17.3% ± 3.5% to 80.7% ± 3.8% (P < 0.01). In the absence of tubulin polymerization or in the presence of high salt, no Ad was found in the pellet. Ad binding to microtubules was not enhanced by bovine brain MAPs enriched for tau protein or by the addition of bovine serum albumin. Enhanced Ad-microtubule binding was also observed by using a fraction of MAPs purified from lung A549 epithelial cell lysate which contained cytoplasmic dynein. Ad-microtubule interaction was sensitive to the addition of ATP, a hallmark of cytoplasmic dynein-dependent microtubule interactions. Immunodepletion of cytoplasmic dynein from the A549 cell lysate abolished the MAP-enhanced Ad-microtubule binding. The interaction of Ad with both dynein and dynactin complexes was demonstrated by coimmunoprecipitation. Partially uncoated capsids isolated from cells 40 min after infection also exhibited microtubule binding. In summary, the primary mode of Ad attachment to microtubules occurs though cytoplasmic dynein-mediated binding.

Abstract 4535: The mTOR complex 2 promotes glioblastoma migration via the interactions with multiple actin-binding and microtubule-associated proteins

2019 ◽  
Author(s):  
Naphat Chantaravisoot ◽  
Piriya Wongkongkathep ◽  
Narawit Pacharakullanon ◽  
Fuyuhiko Tamanoi ◽  
Joseph A. Loo ◽  
...  
Keyword(s):  
Actin Binding ◽  
Microtubule Associated Proteins ◽  
Associated Proteins

Change in distribution of cytoskeleton-associated proteins, lasp-1 and palladin, during uterine receptivity in the rat endometrium

2018 ◽  
Vol 30 (11) ◽  
pp. 1482 ◽  
Author(s):  
Leigh Nicholson ◽  
Laura Lindsay ◽  
Christopher R. Murphy
Keyword(s):  
Plasma Membrane ◽  
Focal Adhesion ◽  
Early Pregnancy ◽  
Morphological Changes ◽  
Cytoskeletal Proteins ◽  
Initial Contact ◽  
Luminal Epithelium ◽  
Uterine Receptivity ◽  
Uterine Lumen ◽  
Associated Proteins

The epithelium of the uterine lumen is the first point of contact with the blastocyst before implantation. To facilitate pregnancy, these uterine epithelial cells (UECs) undergo morphological changes specific to the receptive uterus. These changes include basal, lateral and apical alterations in the plasma membrane of UECs. This study looked at the cytoskeletal and focal adhesion-associated proteins, lasp-1 and palladin, in the uterus during early pregnancy in the rat. Two palladin isoforms, 140 kDa and 90 kDa, were analysed, with the migration-associated 140-kDa isoform increasing significantly at the time of implantation when compared with the time of fertilisation. Lasp-1 was similarly increased at this time, whilst also being located predominantly apically and laterally in the UECs, suggesting a role in the initial contact between the UECs and the blastocyst. This is the first study to investigate palladin and lasp-1 in the uterine luminal epithelium and suggests an importance for these cytoskeletal proteins in the morphological changes the UECs undergo for pregnancy to occur.

scholarly journals A proteomic survey of microtubule-associated proteins in a R402H TUBA1A mutant mouse

PLoS Genetics ◽  
2020 ◽  
Vol 16 (11) ◽  
pp. e1009104
Author(s):  
Ines Leca ◽  
Alexander William Phillips ◽  
Iris Hofer ◽  
Lukas Landler ◽  
Lyubov Ushakova ◽  
...  
Keyword(s):  
Neuronal Migration ◽  
Molecular Mechanisms ◽  
Mutant Mouse ◽  
Critical Role ◽  
Recurrent Mutation ◽  
Intrinsic Defect ◽  
Quantitative Mass Spectrometry ◽  
Microtubule Associated Proteins ◽  
Associated Proteins ◽  
A Cell

Microtubules play a critical role in multiple aspects of neurodevelopment, including the generation, migration and differentiation of neurons. A recurrent mutation (R402H) in the α-tubulin gene TUBA1A is known to cause lissencephaly with cerebellar and striatal phenotypes. Previous work has shown that this mutation does not perturb the chaperone-mediated folding of tubulin heterodimers, which are able to assemble and incorporate into the microtubule lattice. To explore the molecular mechanisms that cause the disease state we generated a new conditional mouse line that recapitulates the R402H variant. We show that heterozygous mutants present with laminar phenotypes in the cortex and hippocampus, as well as a reduction in striatal size and cerebellar abnormalities. We demonstrate that homozygous expression of the R402H allele causes neuronal death and exacerbates a cell intrinsic defect in cortical neuronal migration. Microtubule sedimentation assays coupled with quantitative mass spectrometry demonstrated that the binding and/or levels of multiple microtubule associated proteins (MAPs) are perturbed by the R402H mutation including VAPB, REEP1, EZRIN, PRNP and DYNC1l1/2. Consistent with these data we show that the R402H mutation impairs dynein-mediated transport which is associated with a decoupling of the nucleus to the microtubule organising center. Our data support a model whereby the R402H variant is able to fold and incorporate into microtubules, but acts as a gain of function by perturbing the binding of MAPs.

scholarly journals Reconstitution of dynein transport to the microtubule plus end by kinesin

eLife ◽  
2014 ◽  
Vol 3 ◽  
Author(s):  
Anthony J Roberts ◽  
Brian S Goodman ◽  
Samara L Reck-Peterson
Keyword(s):  
Single Molecule ◽  
Molecular Motor ◽  
Cytoplasmic Dynein ◽  
Directed Motion ◽  
Microtubule Associated Proteins ◽  
Spatial Regulation ◽  
Intracellular Movement ◽  
Associated Proteins ◽  
Directed Motility ◽  
Dynamic Microtubule

Cytoplasmic dynein powers intracellular movement of cargo toward the microtubule minus end. The first step in a variety of dynein transport events is the targeting of dynein to the dynamic microtubule plus end, but the molecular mechanism underlying this spatial regulation is not understood. Here, we reconstitute dynein plus-end transport using purified proteins from S. cerevisiae and dissect the mechanism using single-molecule microscopy. We find that two proteins–homologs of Lis1 and Clip170–are sufficient to couple dynein to Kip2, a plus-end-directed kinesin. Dynein is transported to the plus end by Kip2, but is not a passive passenger, resisting its own plus-end-directed motion. Two microtubule-associated proteins, homologs of Clip170 and EB1, act as processivity factors for Kip2, helping it overcome dynein's intrinsic minus-end-directed motility. This reveals how a minimal system of proteins transports a molecular motor to the start of its track.

Inhibition of neuronal maturation in primary hippocampal neurons from τ deficient mice

2001 ◽  
Vol 114 (6) ◽  
pp. 1179-1187 ◽  
Author(s):  
H.N. Dawson ◽  
A. Ferreira ◽  
M.V. Eyster ◽  
N. Ghoshal ◽  
L.I. Binder ◽  
...  
Keyword(s):  
Hippocampal Neurons ◽  
Axonal Growth ◽  
Neuronal Maturation ◽  
Significant Delay ◽  
Microtubule Associated Proteins ◽  
Conflicting Evidence ◽  
Associated Proteins ◽  
Classic Technique ◽  
Τ Protein ◽  
Deficient Mice

Conflicting evidence supports a role for τ as an essential neuronal cytoskeletal protein or as a redundant protein whose function can be fulfilled by other microtubule-associated proteins. To investigate the function of τ in axonogenesis, we created τ deficient mice by disrupting the TAU gene. The engineered mice do not express the τ protein, appear physically normal and are able to reproduce. In contrast to a previously reported τ knockout mouse, embryonic hippocampal cultures from τ deficient mice show a significant delay in maturation as measured by axonal and neuritic extensions. The classic technique of selectively enhancing axonal growth by growth on laminin substrates failed to restore normal neuronal maturation of τ knockout neurons. By mating human TAU-gene transgenic and τ knockout mice, we reconstituted τ-deficient neurons with human τ proteins and restored a normal pattern of axonal growth and neuronal maturation. The ability of human τ proteins to rescue τ-deficient mouse neurons confirms that τ expression affects the rate of neurite extension.

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