scholarly journals Molecular basis for the interaction between rabies virus phosphoprotein P and the dynein light chain LC8: dissociation of dynein-binding properties and transcriptional functionality of P

2001 ◽  
Vol 82 (11) ◽  
pp. 2691-2696 ◽  
Author(s):  
Nicolas Poisson ◽  
Eleonore Real ◽  
Yves Gaudin ◽  
Marie-Christine Vaney ◽  
Stephen King ◽  
...  
Keyword(s):  
Light Chain ◽  
Neuronal Nitric Oxide Synthase ◽  
Site Directed Mutagenesis ◽  
Dynein Light Chain ◽  
Binding Properties ◽  
Directed Movement ◽  
Virus Transcription ◽  
Oxide Synthase ◽  
Two Hybrid ◽  
Transcription And Replication

The lyssavirus phosphoprotein P is a co-factor of the viral RNA polymerase and plays a central role in virus transcription and replication. It has been shown previously that P interacts with the dynein light chain LC8, which is involved in minus end-directed movement of organelles along microtubules. Co-immunoprecipitation experiments and the two-hybrid system were used to map the LC8-binding site to the sequence 139RSSEDKSTQTTGR151. Site-directed mutagenesis of residues D143 and Q147 to an A residue abolished binding to LC8. The P–LC8 association is not required for virus transcription, since the double mutant was not affected in its transcription ability in a minigenome assay. Based on the crystal structure of LC8 bound to a peptide from neuronal nitric oxide synthase, a model for the complex between the peptide spanning residues 140–150 of P and LC8 is proposed. This model suggests that P binds LC8 in a manner similar to other LC8 cellular partners.

scholarly journals The hDLG-associated protein DAP interacts with dynein light chain and neuronal nitric oxide synthase

2000 ◽  
Vol 5 (11) ◽  
pp. 905-911 ◽  
Author(s):  
Keiko Haraguchi ◽  
Kiyotoshi Satoh ◽  
Hiroyuki Yanai ◽  
Fumihiko Hamada ◽  
Masahiro Kawabuchi ◽  
...  
Keyword(s):  
Nitric Oxide ◽  
Nitric Oxide Synthase ◽  
Light Chain ◽  
Neuronal Nitric Oxide Synthase ◽  
Dynein Light Chain ◽  
Oxide Synthase

scholarly journals Interaction of the Rabies Virus P Protein with the LC8 Dynein Light Chain

2000 ◽  
Vol 74 (21) ◽  
pp. 10212-10216 ◽  
Author(s):  
Hélène Raux ◽  
Anne Flamand ◽  
Danielle Blondel
Keyword(s):  
Rabies Virus ◽  
Light Chain ◽  
Cytoplasmic Dynein ◽  
Dynein Light Chain ◽  
Directed Movement ◽  
Virus Particles ◽  
P Protein ◽  
Infected Cells ◽  
Transcription And Replication

ABSTRACT The rabies virus P protein is involved in viral transcription and replication but its precise function is not clear. We investigated the role of P (CVS strain) by searching for cellular partners by using a two-hybrid screening of a PC12 cDNA library. We isolated a cDNA encoding a 10-kDa dynein light chain (LC8). LC8 is a component of cytoplasmic dynein involved in the minus end-directed movement of organelles along microtubules. We confirmed that this molecule interacts with P by coimmunoprecipitation in infected cells and in cells transfected with a plasmid encoding P protein. LC8 was also detected in virus particles. Series of deletions from the N- and C-terminal ends of P protein were used to map the LC8-binding domain to the central part of P (residues 138 to 172). These results are relevant to speculate that dynein may be involved in the axonal transport of rabies virus along microtubules through neuron cells.

Binding of Dynein Light Chain (PIN) to Neuronal Nitric Oxide Synthase in the Absence of Inhibition

1998 ◽  
Vol 359 (2) ◽  
pp. 297-304 ◽  
Author(s):  
Ignacio Rodrı́guez-Crespo ◽  
Wesley Straub ◽  
Francisco Gavilanes ◽  
Paul R. Ortiz de Montellano
Keyword(s):  
Nitric Oxide ◽  
Nitric Oxide Synthase ◽  
Light Chain ◽  
Neuronal Nitric Oxide Synthase ◽  
Dynein Light Chain ◽  
Oxide Synthase

scholarly journals Dynein Light Chain 1 Regulates Dynamin-mediated F-Actin Assembly during Sperm Individualization in Drosophila

2005 ◽  
Vol 16 (7) ◽  
pp. 3107-3116 ◽  
Author(s):  
Anindya Ghosh-Roy ◽  
Bela S. Desai ◽  
Krishanu Ray
Keyword(s):  
Light Chain ◽  
Cytoplasmic Dynein ◽  
Actin Dynamics ◽  
Dynein Light Chain ◽  
Actin Assembly ◽  
Cellular Functions ◽  
Directed Movement ◽  
Cellular Processes ◽  
Dynactin Complex

Toward the end of spermiogenesis, spermatid nuclei are compacted and the clonally related spermatids individualize to become mature and active sperm. Studies in Drosophila showed that caudal end-directed movement of a microfilament-rich structure, called investment cone, expels the cytoplasmic contents of individual spermatids. F-actin dynamics plays an important role in this process. Here we report that the dynein light chain 1 (DLC1) of Drosophila is involved in two separate cellular processes during sperm individualization. It is enriched around spermatid nuclei during postelongation stages and plays an important role in the dynein-dynactin–dependent rostral retention of the nuclei during this period. In addition, DDLC1 colocalizes with dynamin along investment cones and regulates F-actin assembly at this organelle by retaining dynamin along the cones. Interestingly, we found that this process does not require the other subunits of cytoplasmic dynein-dynactin complex. Altogether, these observations suggest that DLC1 could independently regulate multiple cellular functions and established a novel role of this protein in F-actin assembly in Drosophila.

scholarly journals Ebolavirus VP35 Interacts with the Cytoplasmic Dynein Light Chain 8

2009 ◽  
Vol 83 (13) ◽  
pp. 6952-6956 ◽  
Author(s):  
Toru Kubota ◽  
Mayumi Matsuoka ◽  
Tsung-Hsien Chang ◽  
Mike Bray ◽  
Steven Jones ◽  
...  
Keyword(s):  
Life Cycle ◽  
Light Chain ◽  
Viral Protein ◽  
Cytoplasmic Dynein ◽  
Type I ◽  
Dynein Light Chain ◽  
Yeast Two Hybrid ◽  
Binding Partners ◽  
Mapping Analysis ◽  
Two Hybrid

ABSTRACT The viral protein VP35 of ebolavirus (EBOV) is implicated to have diverse roles in the viral life cycle. We employed a yeast two-hybrid screen to search for VP35 binding partners and identified the cytoplasmic dynein light chain (DLC8) as a protein that interacts with VP35. Mapping analysis unraveled a consensus motif, SQTQT, within VP35 through which VP35 binds to DLC8. The disruption of DLC8 binding does not affect the ability of VP35 to inhibit type I IFN production. Given that VP35 from various EBOV species interacts with DLC8, this interaction may have a role in regulating the EBOV life cycle.

Dynein light chain interacts with NRF-1 and EWG, structurally and functionally related transcription factors from humans and drosophila

2000 ◽  
Vol 113 (23) ◽  
pp. 4263-4273 ◽  
Author(s):  
R.P. Herzig ◽  
U. Andersson ◽  
R.C. Scarpulla
Keyword(s):  
Transcription Factors ◽  
Gene Product ◽  
Light Chain ◽  
Chemical Crosslinking ◽  
Nuclear Staining ◽  
Dynein Light Chain ◽  
Nuclear Respiratory Factor ◽  
Nuclear Respiratory Factor 1 ◽  
Two Hybrid ◽  
Chain Interaction

Nuclear respiratory factor-1 is a transcriptional activator that has been implicated in the nuclear control of respiratory chain expression. Yeast two-hybrid screens were performed to identify proteins that physically interact with nuclear respiratory factor-1. Saturation screening of both mouse embryo and mouse testis libraries yielded 14 independent clones, all of which represented two different isoforms of dynein light chain. In addition to using the two-hybrid method, the specificity of the nuclear respiratory factor-1/dynein light chain interaction was established by chemical crosslinking of the purified native proteins and by co-immunoprecipitation of nuclear respiratory factor-1 and dynein light chain from mammalian cells. Both two-hybrid and chemical crosslinking assays demonstrated that binding of dynein light chain required the first 26 amino acids of nuclear respiratory factor-1. Although dynein light chain is associated with dynein, a cytoplasmic motor molecule, immunolocalizations showed substantial nuclear staining using several different anti-dynein light chain antibodies. Moreover, fluorescence overlays of confocal images established that nuclear respiratory factor-1 and dynein light chain displayed a very similar nuclear staining pattern. The significance of the nuclear respiratory factor-1/dynein light chain interaction was investigated further by determining whether a similar interaction was conserved between dynein light chain and the erect wing gene product of Drosophila, a protein related to nuclear respiratory factor-1 through its DNA binding domain. Here, we establish that the erect wing gene product can bind and trans-activate transcription through authentic nuclear respiratory factor-1 binding sites. Moreover, the erect wing gene product, like nuclear respiratory factor-1, interacted specifically with dynein light chain both in vitro and in transfected cells. Thus, the interaction with dynein light chain is conserved between transcription factors that are structurally and functionally similar between humans and Drosophila.

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