Identification of novel cellular proteins that bind to the LC8 dynein light chain using a pepscan technique

Rodrı́guez-Crespo, Ignacio; Yelamos, Belèn; Roncal, Fernando; Albar, Juan Pablo; Montellano, Paul R. Ortiz de; Gavilanes, Francisco

doi:10.1016/s0014-5793(01)02718-1

Cited by 95 publications

(108 citation statements)

References 26 publications

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“…The first HSV-1 protein shown to bind to dynein was pUL34, which interacted in vitro with an Nterminal fragment of cytoplasmic dynein 1 intermediate chain (DYNC1I)1a [52]. The HSV-1 viral helicase pUL9 contains the typical LC8-binding motif KSTQT [53], and was recently shown to bind to LC8 in vitro [54]. Despite these findings, pUL34 and pUL9 are unlikely to play significant roles during retrograde transport of HSV-1.…”

Section: Dynein Cofactor: Dynactinmentioning

confidence: 99%

Transport and egress of herpes simplex virus in neurons

Diefenbach

Miranda-Saksena

Douglas

et al. 2007

Reviews in Medical Virology

171

150

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The mechanisms of axonal transport of the alphaherpesviruses, HSV and pseudorabies virus (PrV), in neuronal axons are of fundamental interest, particularly in comparison with other viruses, and offer potential sites for antiviral intervention or development of gene therapy vectors. These herpesviruses are transported rapidly along microtubules (MTs) in the retrograde direction from the axon terminus to the dorsal root ganglion and then anterogradely in the opposite direction. Retrograde transport follows fusion and deenvelopment of the viral capsid at the axonal membrane followed by loss of most of the tegument proteins and then binding of the capsid via one or more viral proteins (VPs) to the retrograde molecular motor dynein. The HSV capsid protein pUL35 has been shown to bind to the dynein light chain Tctex1 but is likely to be accompanied by additional dynein binding of an inner tegument protein. The mechanism of anterograde transport is much more controversial with different processes being claimed for PrV and HSV: separate transport of HSV capsid/tegument and glycoproteins versus PrV transport as an enveloped virion. The controversy has not been resolved despite application, in several laboratories, of confocal microscopy (CFM), realtime fluorescence with viruses dual labelled on capsid and glycoprotein, electron microscopy in situ and immunoelectron microscopy. Different processes for each virus seem counterintuitive although they are the most divergent in the alphaherpesvirus subfamily. Current hypotheses suggest that unenveloped HSV capsids complete assembly in the axonal growth cones and varicosities, whereas with PrV unenveloped capsids are only found travelling in a retrograde direction.

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Section: Dynein Cofactor: Dynactinmentioning

confidence: 99%

Transport and egress of herpes simplex virus in neurons

Diefenbach

Miranda-Saksena

Douglas

et al. 2007

Reviews in Medical Virology

171

150

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“…Rather, it might function as part of the neuronal machinery serving the axonal transport of NOS-I, as it was shown to be part of the microtubule-associated motor protein dynein complexes and hence termed dynein light chain of 8 kDa (LC8, DLC1). It is thought to link the dynein complex to cargo molecules including NOS-I (Rodriguez-Crespo et al, 2001), DLGAP1, GluN3A and PSD-95 (Navarro-Lerida et al, 2004) supporting the notion that DYNLL1 serves as a transport adaptor vehicle for proteins which constitute the glutamatergic postsynaptic complex. This is underscored by data showing that DYNLL1 is also part of the above mentioned NMDA -PSD-95 -NOS-I -DLGAP1 complex, probably trafficking this complex along microtubules and actin cytoskeleton (Haraguchi et al, 2000).…”

Section: Introductionmentioning

confidence: 99%

The genetic contribution of the NO system at the glutamatergic post-synapse to schizophrenia: Further evidence and meta-analysis

Weber

Klamer

Freudenberg

et al. 2014

European Neuropsychopharmacology

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NO is a pleiotropic signaling molecule and has an important role for cognition and emotion. In the brain, NO is produced by neuronal nitric oxide synthase (NOS-I, encoded by NOS1) coupled to the NMDA receptor via PDZ interactions; this protein-protein interaction is disrupted upon binding of NOS1 adaptor protein (encoded by NOS1AP) to NOS-I. As both NOS1 and NOS1AP were associated with schizophrenia, we here investigated these genes in greater detail by genotyping new samples and conducting a meta-analysis of our own and published data. In doing so, we confirmed association of both genes with schizophrenia and found evidence for their interaction in increasing risk towards disease. Our strongest finding was the NOS1 promoter SNP rs41279104, yielding an odds ratios of 1.29 in the metaanalysis. As findings from heterologous cell systems have suggested that the risk allele decreases gene expression, we studied the effect of the variant on NOS1 expression in human post-mortem brain samples and found that the risk allele significantly decreases expression of NOS1 in the prefrontal cortex. Bioinformatic analyses suggest that this might be due the replacement of six transcription factor binding sites by two new binding sites as a consequence of proxy SNPs. Taken together, our data argue that genetic variance in NOS1 resulting in lower prefrontal brain expression of this gene contributes to schizophrenia liability, and that NOS1 interacts with NOS1AP in doing so. The NOS1-NOS1AP PDZ interface may thus well constitute a novel target for small molecules in at least some forms of schizophrenia.

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“…16 Additionally, pepscan techniques have demonstrated the ability of short peptides containing these consensus motifs to bind recombinant LC8 in vitro. [17][18][19] Due to the identification of consensus amino acid motifs that bind LC8, this subunit is considered an attractive target for linking cargo to the dynein motor complex via a short peptide.…”

Section: Introductionmentioning

confidence: 99%

Evaluation of an LC8-Binding Peptide for the Attachment of Artificial Cargo to Dynein

Bergen

Pun

2006

Mol. Pharmaceutics

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The limited cytoplasmic mobility of non-viral gene carriers is likely to contribute to their low transfection efficiency. This limitation could be overcome by mimicking the viral strategy of recruiting the dynein motor complex for efficient transport toward the host cell nucleus. A promising approach for attaching artificial cargo to dynein is through an adaptor peptide that binds the 8-kDa light chain (LC8) found in the cargo-binding region of the dynein complex. Several viral proteins that bind LC8 have in common an LC8-binding motif defined by (K/R)XTQT. Short peptides containing this motif have also been shown to bind recombinant LC8 in vitro. However, since the majority of intracellular LC8 exists outside of the dynein complex, it remains unclear whether peptides displaying this LC8-binding motif can access and bind to dynein-associated LC8. In this study, we employed biochemical analysis to investigate the feasibility of attaching artificial cargo to the dynein motor complex using a peptide displaying the well-characterized LC8-binding motif. We report that free intracellular LC8 bound specifically to an LC8-binding (TQT) peptide and not to a control peptide with a mutated LC8-binding motif. However, a similar binding interaction between the TQT peptide and intracellular dynein was not detected. To determine whether dynein binding of the TQT peptide was prevented by competition with free intracellular LC8 or due to the inability of the peptide to access its LC8 binding site in the dynein complex, the TQT peptide was evaluated for its ability to bind either purified LC8 or purified dynein. Our results demonstrate that, while the TQT peptide readily binds free LC8, it cannot bind to dynein-associated LC8. The results emphasize the need to identify functional dynein-binding peptides and highlight the importance of designing peptides that bind to the intact dynein motor complex.

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Identification of novel cellular proteins that bind to the LC8 dynein light chain using a pepscan technique

Cited by 95 publications

References 26 publications

Transport and egress of herpes simplex virus in neurons

Transport and egress of herpes simplex virus in neurons

The genetic contribution of the NO system at the glutamatergic post-synapse to schizophrenia: Further evidence and meta-analysis

Evaluation of an LC8-Binding Peptide for the Attachment of Artificial Cargo to Dynein

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