NudC associates with Lis1 and the dynein motor at the leading pole of neurons (original) (raw)
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Current Biology, 2001
Cytoplasmic dynein is a multisubunit, minus end-Results and discussion directed microtubule motor that uses dynactin as Both NUDF (the LIS1-like protein) and NUDA (the cytoplasmic dynein heavy chain) an accessory complex to perform various in vivo localize to microtubule ends functions including vesicle transport, spindle To investigate the in vivo behavior of the LIS1-homoloassembly, and nuclear distribution [1]. We previously gous protein, NUDF, we constructed a strain expressing showed that in the filamentous fungus Aspergillus GFP-NUDF, which allowed us to observe NUDF localnidulans, a GFP-tagged cytoplasmic dynein heavy ization in living cells. In this strain, the GFP tag is at chain (NUDA) forms comet-like structures that the N terminus of NUDF, and the GFP-NUDF fusion exhibited microtubule-dependent movement toward protein is functional (see the Supplementary material and back from the hyphal tip [2]. Here we available with this article on the internet). Interestingly, demonstrate that another protein in the NUDA GFP-NUDF forms comet-like structures that migrate rappathway, NUDF, which is homologous to the idly toward the hyphal tip (Figure 1). Retrograde movehuman LIS1 protein involved in brain development ments of GFP-NUDF comets away from the hyphal tip
Current Biology, 1998
Important clues to how the mammalian cerebral cortex develops are provided by the analysis of genetic diseases that cause cortical malformations [1-5]. People with Miller-Dieker syndrome (MDS) or isolated lissencephaly sequence (ILS) have a hemizygous deletion or mutation in the LIS1 gene [3,6]; both conditions are characterized by a smooth cerebral surface, a thickened cortex with four abnormal layers, and misplaced neurons [7,8]. LIS1 is highly expressed in the ventricular zone and the cortical plate [9,10], and its product, Lis1, has seven WD repeats [3]; several proteins with such repeats have been shown to interact with other polypeptides, giving rise to multiprotein complexes [11]. Lis1 copurifies with platelet-activating factor acetylhydrolase subunits a1 and a2 [12], and with tubulin; it also reduces microtubule catastrophe events in vitro [13]
The Nuclear Migration Protein NUDF/LIS1 Forms a Complex with NUDC and BNFA at Spindle Pole Bodies
Eukaryotic Cell, 2008
Nuclear migration depends on microtubules, the dynein motor complex, and regulatory components like LIS1 and NUDC. We sought to identify new binding partners of the fungal LIS1 homolog NUDF to clarify its function in dynein regulation. We therefore analyzed the association between NUDF and NUDC in Aspergillus nidulans. NUDF and NUDC directly interacted in yeast two-hybrid experiments via NUDF's WD40 domain. NUDC-green fluorescent protein (NUDC-GFP) was localized to immobile dots in the cytoplasm and at the hyphal cortex, some of which were spindle pole bodies (SPBs). We showed by bimolecular fluorescence complementation microscopy that NUDC directly interacted with NUDF at SPBs at different stages of the cell cycle. Applying tandem affinity purification, we isolated the NUDF-associated protein BNFA (for binding to NUDF). BNFA was dispensable for growth and for nuclear migration. GFP-BNFA fusions localized to SPBs at different stages of the cell cycle. This localization depended on NUDF, since the loss of NUDF resulted in the cytoplasmic accumulation of BNFA. BNFA did not bind to NUDC in a yeast two-hybrid assay. These results show that the conserved NUDF and NUDC proteins play a concerted role at SPBs at different stages of the cell cycle and that NUDF recruits additional proteins specifically to the dynein complex at SPBs.
Nuclear migration, nucleokinesis and lissencephaly
Trends in Cell Biology, 1998
During the past 20 years, biologists have become used to finding that proteins first identified in simple, genetically manipulable eukaryotic organisms are conserved in higher eukaryotes. This article draws attention to the similarity between NUDF protein, which is required for nuclear migration in the filamentous fungus Aspergillus nidulans, and a mammalian homologue, LIS1, whose malfunction causes lissencephaly, a neuronal migration disease. The authors suggest that there might be an underlying similarity of mechanism between nuclear migration in the fungus and neuronal migration in the brain.
Nuclear migration: Cortical anchors for cytoplasmic dynein
2001
The nucleus is not just a passive body that rolls around at random inside the sack of a eukaryotic cell. Controlled nuclear movements are important in a number of contexts -for example, during very early Drosophila development, where they play a key role in establishing oocyte polarity, and in the yeast Saccharomyces cerevisiae, where they are required during budding. Nuclear migration in budding yeast was first proposed by Hartwell et al.
Journal of neurocytology, 1998
Neurons are terminally post-mitotic cells that utilize their microtubule arrays for the growth and maintenance of axons and dendrites rather than for the formation of mitotic spindles. Recent studies from our laboratory suggest that the mechanisms that organize the axonal and dendritic microtubule arrays may be variations on the same mechanisms that organize the mitotic spindle in dividing cells. In particular, we have identified molecular motor proteins that serve analogous functions in the establishment of these seemingly very different microtubule arrays. In the present study, we have sought to determine whether a non-motor protein termed NuMA is also a component of both systems. NuMA is a approximately 230 kDa structural protein that is present exclusively in the nucleus during interphase. During mitosis, NuMA forms aggregates that interact with microtubules and certain motor proteins. As a result of these interactions, NuMA is thought to draw together the minus-ends of microtub...
Cell Cycle, 2014
Cytoplasmic dynein is recruited to the cell cortex in early mitosis, where it can generate pulling forces on astral microtubules to position the mitotic spindle. Recent work has shown that dynein displays a dynamic asymmetric cortical localization, and that dynein recruitment is negatively regulated by spindle pole-proximity. this results in oscillating dynein recruitment to opposite sides of the cortex to center the mitotic spindle. However, although the centrosome-derived signal that promotes displacement of dynein has been identified, it is currently unknown how dynein is re-recruited to the cortex once it has been displaced. Here we show that re-recruitment of cortical dynein requires astral microtubules. We find that microtubules are necessary for the sustained localized enrichment of dynein at the cortex. Furthermore, we show that stabilization of astral microtubules causes spindle misorientation, followed by mispositioning of dynein at the cortex. thus, our results demonstrate the importance of astral microtubules in the dynamic regulation of cortical dynein recruitment in mitosis.
Molecular Biology of the Cell, 2001
During meiotic prophase in fission yeast, the nucleus migrates back and forth between the two ends of the cell, led by the spindle pole body (SPB). This nuclear oscillation is dependent on astral microtubules radiating from the SPB and a microtubule motor, cytoplasmic dynein. Here we have examined the dynamic behavior of astral microtubules labeled with the green fluorescent protein during meiotic prophase with the use of optical sectioning microscopy. During nuclear migrations, the SPB mostly follows the microtubules that extend toward the cell cortex. SPB migrations start when these microtubules interact with the cortex and stop when they disappear, suggesting that these microtubules drive nuclear migrations. The microtubules that are followed by the SPB often slide along the cortex and are shortened by disassembly at their ends proximal to the cortex. In dynein-mutant cells, where nuclear oscillations are absent, the SPB never migrates by following microtubules, and microtubule assembly/disassembly dynamics is significantly altered. Based on these observations, together with the frequent accumulation of dynein at a cortical site where the directing microtubules interact, we propose a model in which dynein drives nuclear oscillation by mediating cortical microtubule interactions and regulating the dynamics of microtubule disassembly at the cortex. ‡ Corresponding author. E-mail address: ayumu@crl.go.jp. Abbreviations used: DHC, dynein heavy chain; GFP, green fluorescent protein; SPB, spindle pole body.