scholarly journals Interactions of Cytoplasmic Dynein Light Chains Tctex-1 and LC8 with the Intermediate Chain IC74†

Biochemistry ◽  
2002 ◽  
Vol 41 (13) ◽  
pp. 4302-4311 ◽  
Author(s):  
Moses Makokha ◽  
Michael Hare ◽  
Mingang Li ◽  
Thomas Hays ◽  
Elisar Barbar
Keyword(s):  
Cytoplasmic Dynein ◽  
Light Chains ◽  
Dynein Light Chains ◽  
Intermediate Chain

scholarly journals Interactions of LC8 with N-Terminal Segments of the Intermediate Chain of Cytoplasmic Dynein

2003 ◽  
Vol 3 ◽  
pp. 647-654 ◽  
Author(s):  
Afua Nyarko ◽  
Michael Hare ◽  
Moses Makokha ◽  
Elisar Barbar
Keyword(s):  
Limited Proteolysis ◽  
Coiled Coil ◽  
Cytoplasmic Dynein ◽  
Light Chains ◽  
Ordered Structure ◽  
Dynein Motor ◽  
Dynein Light Chains ◽  
Significant Conformational Change ◽  
Intermediate Chain

LC8, a highly conserved 10-kDa light chain, and IC74, a 74-kDa intermediate chain, are presumed to promote the assembly of the cytoplasmic dynein motor protein complex and to be engaged in the controlled binding and release of cargo. The interactions of LC8 from Drosophila melanogaster with constructs of IC74 were characterized in vitro by affinity methods, limited proteolysis, and circular dichroism spectroscopy. Previously, we have performed limited proteolysis on the N-terminal domain of IC74, IC(1-289), when free and when bound to dynein light chains LC8 and Tctex-1[1]. We have also shown that upon addition of LC8, IC(1-289) undergoes a significant conformational change from a largely unfolded to a more ordered structure. The purpose of the work presented here is to determine whether residues 1-30 in IC74, predicted to be in a coiled coil, are involved in the stabilization of the protein upon binding to LC8. Constructs of IC74, IC(1-143), and IC(30-143) that include the LC8 binding site but with and without the first 30 residues were prepared, and their binding and protection patterns were compared to our previous results for IC(1-289). The results suggest that coiled coil residues 1-30 are not responsible for the increase in structure we observe when IC(1-289) binds to LC8.

scholarly journals Somatic CRISPR–Cas9-induced mutations reveal roles of embryonically essential dynein chains in Caenorhabditis elegans cilia

2015 ◽  
Vol 208 (6) ◽  
pp. 683-692 ◽  
Author(s):  
Wenjing Li ◽  
Peishan Yi ◽  
Guangshuo Ou
Keyword(s):  
Caenorhabditis Elegans ◽  
Intraflagellar Transport ◽  
Cytoplasmic Dynein ◽  
Regulatory Mechanisms ◽  
Light Chains ◽  
Induced Mutations ◽  
Functional Studies ◽  
Cilium Formation ◽  
Dynein Intermediate Chain ◽  
Intermediate Chain

Cilium formation and maintenance require intraflagellar transport (IFT). Although much is known about kinesin-2–driven anterograde IFT, the composition and regulation of retrograde IFT-specific dynein remain elusive. Components of cytoplasmic dynein may participate in IFT; however, their essential roles in cell division preclude functional studies in postmitotic cilia. Here, we report that inducible expression of the clustered regularly interspaced short palindromic repeats (CRISPR)–Cas9 system in Caenorhabditis elegans generated conditional mutations in IFT motors and particles, recapitulating ciliary defects in their null mutants. Using this method to bypass the embryonic requirement, we show the following: the dynein intermediate chain, light chain LC8, and lissencephaly-1 regulate retrograde IFT; the dynein light intermediate chain functions in dendrites and indirectly contributes to ciliogenesis; and the Tctex and Roadblock light chains are dispensable for cilium assembly. Furthermore, we demonstrate that these components undergo biphasic IFT with distinct transport frequencies and turnaround behaviors. Together, our results suggest that IFT–dynein and cytoplasmic dynein have unique compositions but also share components and regulatory mechanisms.

scholarly journals Localization of Cytoplasmic Dynein Light-intermediate Chain mRNA in the Rat Testis Using In Situ Hybridization.

1998 ◽  
Vol 23 (1) ◽  
pp. 9-15 ◽  
Author(s):  
Seishi Maeda ◽  
Sang-Yoon Nam ◽  
Masahiko Fujisawa ◽  
Nobuaki Nakamuta ◽  
Kenji Ogawa ◽  
...  
Keyword(s):  
In Situ Hybridization ◽  
Cytoplasmic Dynein ◽  
Rat Testis ◽  
Intermediate Chain

scholarly journals Structure of Tctex-1 and Its Interaction with Cytoplasmic Dynein Intermediate Chain

2001 ◽  
Vol 276 (17) ◽  
pp. 14067-14074 ◽  
Author(s):  
Yu-Keung Mok ◽  
Kevin W.-H. Lo ◽  
Mingjie Zhang
Keyword(s):  
Cytoplasmic Dynein ◽  
Dynein Intermediate Chain ◽  
Intermediate Chain

scholarly journals PLAC-24 Is a Cytoplasmic Dynein-Binding Protein That Is Recruited to Sites of Cell-Cell Contact

2002 ◽  
Vol 13 (5) ◽  
pp. 1722-1734 ◽  
Author(s):  
Sher Karki ◽  
Lee A. Ligon ◽  
Jamison DeSantis ◽  
Mariko Tokito ◽  
Erika L. F. Holzbaur
Keyword(s):  
Epithelial Cells ◽  
Recent Observation ◽  
Polyclonal Antibodies ◽  
Adherens Junction ◽  
Cytoplasmic Dynein ◽  
Cell Contact ◽  
Dynein Intermediate Chain ◽  
Cellular Cytoskeleton ◽  
Cell Cell ◽  
Intermediate Chain

We screened for polypeptides that interact specifically with dynein and identified a novel 24-kDa protein (PLAC-24) that binds directly to dynein intermediate chain (DIC). PLAC-24 is not a dynactin subunit, and the binding of PLAC-24 to the dynein intermediate chain is independent of the association between dynein and dynactin. Immunocytochemistry using PLAC-24–specific polyclonal antibodies revealed a punctate perinuclear distribution of the polypeptide in fibroblasts and isolated epithelial cells. However, as epithelial cells in culture make contact with adjacent cells, PLAC-24 is specifically recruited to the cortex at sites of contact, where the protein colocalizes with components of the adherens junction. Disruption of the cellular cytoskeleton with latrunculin or nocodazole indicates that the localization of PLAC-24 to the cortex is dependent on intact actin filaments but not on microtubules. Overexpression of β-catenin also leads to a loss of PLAC-24 from sites of cell-cell contact. On the basis of these data and the recent observation that cytoplasmic dynein is also localized to sites of cell-cell contact in epithelial cells, we propose that PLAC-24 is part of a multiprotein complex localized to sites of intercellular contact that may function to tether microtubule plus ends to the actin-rich cellular cortex.

scholarly journals The Herpes Simplex Virus 1 UL34 Protein Interacts with a Cytoplasmic Dynein Intermediate Chain and Targets Nuclear Membrane

2000 ◽  
Vol 74 (3) ◽  
pp. 1355-1363 ◽  
Author(s):  
Guo-Jie Ye ◽  
Kevin T. Vaughan ◽  
Richard B. Vallee ◽  
Bernard Roizman
Keyword(s):  
Herpes Simplex Virus ◽  
Herpes Simplex ◽  
Nuclear Membrane ◽  
Viral Proteins ◽  
Cytoplasmic Dynein ◽  
Dependent Manner ◽  
Viral Dna ◽  
Herpes Simplex Virus 1 ◽  
Simplex Virus ◽  
Intermediate Chain

ABSTRACT To express the function encoded in its genome, the herpes simplex virus 1 capsid-tegument structure released by deenvelopment during entry into cells must be transported retrograde to the nuclear pore where viral DNA is released into the nucleus. This path is essential in the case of virus entering axons of dorsal root ganglia. The objective of the study was to identify the viral proteins that may be involved in the transport. We report the following findings. (i) The neuronal isoform of the intermediate chain (IC-1a) of the dynein complex pulled down, from lysates of [35S]methionine-labeled infected cells, two viral proteins identified as the products of UL34 and UL31 open reading frames, respectively. UL34 protein is a virion protein associated with cellular membranes and phosphorylated by the viral kinase US3. UL31 protein is a largely insoluble, evenly dispersed nuclear phosphoprotein required for optimal processing and packaging of viral DNA into preformed capsids. Reciprocal pulldown experiments verified the interaction of IC-1a and UL34 protein. In similar experiments, UL34 protein was found to interact with UL31 protein and the major capsid protein ICP5. (ii) To determine whether UL34 protein is transported to the nuclear membrane, a requirement if it is involved in transport, the UL34 protein was inserted into a baculovirus vector under the cytomegalovirus major early promoter. Cells infected with the recombinant baculovirus expressed UL34 protein in a dose-dependent manner, and the UL34 protein localized primarily in the nuclear membrane. An unexpected finding was that UL34-expressing cells showed a dissociation of the inner and outer nuclear membranes reminiscent of the morphologic changes seen in cells productively infected with herpes simplex virus 1. UL34, like many other viral proteins, may have multiple functions expressed both early and late in infection.

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