Dynein light chain regulates axonal trafficking and synaptic levels of Bassoon

Fejtová, Anna; Davydova, Daria; Bischof, Ferdinand; Lazarevic, Vesna; Altrock, Wilko D.; Romorini, Stefano; Schöne, Cornelia; Zuschratter, Werner; Kreutz, Michael R.; Garner, Craig C.; Ziv, Noam; Gundelfinger, Eckart D.

doi:10.1083/jcb.200807155

Cited by 92 publications

(96 citation statements)

References 63 publications

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“…Higher apparent affinity of DYNLL binding to dimeric protein fragments was first noted by studying DYNLL2 and myoVa interactions (2) and later attributed to avidity (15). This hypothesis questioned the original proposal of heterodimeric cargo adapter role of DYNLL on dynein and myoVa motor complexes (6,7,21,(55)(56)(57)(58).…”

Section: Fine-tuning Of Affinities Is Achieved Through Diversity Of Bmentioning

confidence: 85%

Affinity, Avidity, and Kinetics of Target Sequence Binding to LC8 Dynein Light Chain Isoforms

Radnai

Rapali

Hódi

et al. 2010

Journal of Biological Chemistry

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LC8 dynein light chain (DYNLL) is a highly conserved eukaryotic hub protein with dozens of binding partners and various functions beyond being a subunit of dynein and myosin Va motor proteins. Here, we compared the kinetic and thermodynamic parameters of binding of both mammalian isoforms, DYNLL1 and DYNLL2, to two putative consensus binding motifs (KXTQTX and XG(I/V)QVD) and report only subtle differences. Peptides containing either of the above motifs bind to DYNLL2 with micromolar affinity, whereas a myosin Va peptide (lacking the conserved Gln) and the noncanonical Pak1 peptide bind with K d values of 9 and 40 M, respectively. Binding of the KXTQTX motif is enthalpy-driven, although that of all other peptides is both enthalpy-and entropy-driven. Moreover, the KXTQTX motif shows strikingly slower off-rate constant than the other motifs. As most DYNLL partners are homodimeric, we also assessed the binding of bivalent ligands to DYNLL2. Compared with monovalent ligands, a significant avidity effect was found as follows: K d values of 37 and 3.5 nM for a dimeric myosin Va fragment and a Leu zipper dimerized KXTQTX motif, respectively. Ligand binding kinetics of DYNLL can best be described by a conformational selection model consisting of a slow isomerization and a rapid binding step. We also studied the binding of the phosphomimetic S88E mutant of DYNLL2 to the dimeric myosin Va fragment, and we found a significantly lower apparent K d value (3 M). We conclude that the thermodynamic and kinetic fine-tuning of binding of various ligands to DYNLL could have physiological relevance in its interaction network.

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Section: Fine-tuning Of Affinities Is Achieved Through Diversity Of Bmentioning

confidence: 85%

Affinity, Avidity, and Kinetics of Target Sequence Binding to LC8 Dynein Light Chain Isoforms

Radnai

Rapali

Hódi

et al. 2010

Journal of Biological Chemistry

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“…Its crucial role has been demonstrated in several organisms, including Drosophila, in which null mutations result in embryonic lethality (Dick et al, 1996). DLC associates with a wide variety of proteins, including neuronal nitric oxide synthase (nNOS) (Jaffrey and Snyder, 1996), gephyrin (Fuhrmann et al, 2002), bassoon (Fejtova et al, 2009), the proapoptotic factor Bim (Puthalakath et al, 1999), transcriptional factors Swallow (Schnorrer et al, 2000) and Trps1 (Kaiser et al, 2003), viral proteins (Raux et al, 2000) and p53 binding protein 1 (Lo et al, 2005). Because DLC is a light chain of motor proteins, its role in trafficking has been widely assessed.…”

Section: Introductionmentioning

confidence: 99%

GKAP-DLC2 interaction organizes postsynaptic scaffold complex to enhance synaptic NMDA receptor activity

Moutin

Raynaud

Fagni

et al. 2012

Journal of Cell Science

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SummaryAt glutamatergic brain synapses, scaffolding proteins regulate receptor location and function. The targeting and organization of scaffolding proteins in the postsynaptic density (PSD) is poorly understood, but it is known that a core protein of the glutamatergic receptor postsynaptic scaffold complex, guanylate-kinase-associated protein (GKAP) interacts with dynein light chain 2 (DLC2, also known as DYNLL2), a protein associated with molecular motors. In the present study, we combined BRET imaging, immunostaining and electrophysiological recording to assess the role of the GKAP-DLC2 interaction in the functional organization of the glutamatergic synapse. We found that GKAP-DLC2 interaction in dendritic spine stabilizes scaffolding protein expression at the PSD and enhances synaptic NMDA receptor activity. Moreover, the GKAP-DLC2 functional interaction is favored by sustained synaptic activity. These data identify a regulatory pathway of synaptic transmission that depends on activity-induced remodelling of the postsynaptic scaffold protein complex.

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“…GKAP also interacts with the dynein light chain [DLC, also known as LC8 and DYNLL (Naisbitt et al, 2000)]. DLC is an essential component of the dynein and myosin V molecular motors (Benashski et al, 1997), but is also found in numerous other complexes (Fejtova et al, 2009;Fuhrmann et al, 2002;Jaffrey and Snyder, 1996;Lo et al, 2005;Puthalakath et al, 1999;Raux et al, 2000;Schnorrer et al, 2000). Thus, in addition to its role in transport as a molecular motor light chain (Lee et al, 2006;Navarro et al, 2004;Schnorrer et al, 2000), DLC could be an ordered hub protein as well, promoting the oligomerization and ordering of the natively disordered monomeric proteins with which it interacts (Benison et al, 2006;Nyarko et al, 2004;Wang et al, 2004).…”

Section: Introductionmentioning

confidence: 99%

Stoichiometry of scaffold complexes in living neurons - DLC2 as a dimerization engine for GKAP

Moutin

Compan

Raynaud

et al. 2014

Journal of Cell Science

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Quantitative spatio-temporal characterization of protein interactions in living cells remains a major challenge facing modern biology. We have investigated in living neurons the spatial dependence of the stoichiometry of interactions between two core proteins of the N-methyl-D-aspartate (NMDA)-receptor-associated scaffolding complex, GKAP (also known as DLGAP1) and DLC2 (also known as DYNLL2), using a novel variation of fluorescence fluctuation microscopy called two-photon scanning number and brightness (sN&B). We found that dimerization of DLC2 was required for its interaction with GKAP, which, in turn, potentiated GKAP selfassociation. In the dendritic shaft, the DLC2-GKAP heterooligomeric complexes were composed mainly of two DLC2 and two GKAP monomers, whereas, in spines, the hetero-complexes were much larger, with an average of ,16 DLC2 and ,13 GKAP monomers. Disruption of the GKAP-DLC2 interaction strongly destabilized the oligomers, decreasing the spine-preferential localization of GKAP and inhibiting NMDA receptor activity. Hence, DLC2 serves a hub function in the control of glutamatergic transmission by ordering GKAP-containing complexes in dendritic spines. Beyond illuminating the role of DLC2-GKAP interactions in glutamatergic signaling, these data underscore the power of the sN&B approach for quantitative spatio-temporal imaging of other important protein complexes.

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Dynein light chain regulates axonal trafficking and synaptic levels of Bassoon

Cited by 92 publications

References 63 publications

Affinity, Avidity, and Kinetics of Target Sequence Binding to LC8 Dynein Light Chain Isoforms

Affinity, Avidity, and Kinetics of Target Sequence Binding to LC8 Dynein Light Chain Isoforms

GKAP-DLC2 interaction organizes postsynaptic scaffold complex to enhance synaptic NMDA receptor activity

Stoichiometry of scaffold complexes in living neurons - DLC2 as a dimerization engine for GKAP

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