Abstract:Metazoan dynein moves processively with the aid of dynactin and the endosomal cargo adaptor Hook3. A structure–function study of Hook3 reveals how it assembles dynein with dynactin and suggests that an additional step of allosteric activation is required beyond complex assembly.
“…NIN was identified in the IC1 and IC2 BioID datasets, NINL and daple in the IC2 BioID dataset, and girdin in the LIC1 dataset (Supplementary file 1). Some construct optimization was necessary to determine the dynein/dynactin interacting region of each candidate activator and we used the literature to guide this process (Casenghi et al, 2005; Schroeder and Vale, 2016) (Figure 4C and D and Figure 4—figure supplement 2A). 10.7554/eLife.28257.008Figure 4.A secondary screen identifies candidate activators of dynein/dynactin motility.( A ) A schematic of our secondary screen.…”
In human cells, cytoplasmic dynein-1 is essential for long-distance transport of many cargos, including organelles, RNAs, proteins, and viruses, towards microtubule minus ends. To understand how a single motor achieves cargo specificity, we identified the human dynein interactome by attaching a promiscuous biotin ligase (‘BioID’) to seven components of the dynein machinery, including a subunit of the essential cofactor dynactin. This method reported spatial information about the large cytosolic dynein/dynactin complex in living cells. To achieve maximal motile activity and to bind its cargos, human dynein/dynactin requires ‘activators’, of which only five have been described. We developed methods to identify new activators in our BioID data, and discovered that ninein and ninein-like are a new family of dynein activators. Analysis of the protein interactomes for six activators, including ninein and ninein-like, suggests that each dynein activator has multiple cargos.DOI:
http://dx.doi.org/10.7554/eLife.28257.001
“…NIN was identified in the IC1 and IC2 BioID datasets, NINL and daple in the IC2 BioID dataset, and girdin in the LIC1 dataset (Supplementary file 1). Some construct optimization was necessary to determine the dynein/dynactin interacting region of each candidate activator and we used the literature to guide this process (Casenghi et al, 2005; Schroeder and Vale, 2016) (Figure 4C and D and Figure 4—figure supplement 2A). 10.7554/eLife.28257.008Figure 4.A secondary screen identifies candidate activators of dynein/dynactin motility.( A ) A schematic of our secondary screen.…”
In human cells, cytoplasmic dynein-1 is essential for long-distance transport of many cargos, including organelles, RNAs, proteins, and viruses, towards microtubule minus ends. To understand how a single motor achieves cargo specificity, we identified the human dynein interactome by attaching a promiscuous biotin ligase (‘BioID’) to seven components of the dynein machinery, including a subunit of the essential cofactor dynactin. This method reported spatial information about the large cytosolic dynein/dynactin complex in living cells. To achieve maximal motile activity and to bind its cargos, human dynein/dynactin requires ‘activators’, of which only five have been described. We developed methods to identify new activators in our BioID data, and discovered that ninein and ninein-like are a new family of dynein activators. Analysis of the protein interactomes for six activators, including ninein and ninein-like, suggests that each dynein activator has multiple cargos.DOI:
http://dx.doi.org/10.7554/eLife.28257.001
“…, 2014; Olenick et al. , 2016; Schroeder and Vale, 2016). Of note, sequence searches could not identify A. nidulans homologues of the coiled-coil–containing prototypic mammalian RAB11 effector and dynein adaptor FIP3 (Horgan et al.…”
In Aspergillus nidulans, the distribution of exocytic carriers involves interplay between kinesin-1, myosin-5, and dynein. Engagement of the dynein complex to these carriers requires dynactin p25, but, unlike that of early endosomes, it does not require the Hook complex.
Mechanosensory transduction for senses such as proprioception, touch, balance, acceleration, hearing and pain relies on mechanotransduction channels, which convert mechanical stimuli into electrical signals in specialized sensory cells1. How force gates mechanotransduction channels is a central question in the field, for which there are two major models. One is the membrane-tension model: force applied to the membrane generates a change in membrane tension that is sufficient to gate the channel, as in the case of bacterial MscL channel and certain eukaryotic potassium channels2-5. The other is the tether model: force is transmitted via a tether to gate the channel. Recent study suggests that NOMPC, a mechanotransduction channel that mediates hearing and touch sensation in Drosophila, is gated by tethering of its ankyrin repeat (AR) domain to microtubules of the cytoskeleton6. Thus, a goal of studying NOMPC is to reveal the underlying mechanism of force induced gating, which could serve as a paradigm of the tether model. NOMPC, a Transient Receptor Potential (TRP) channel and the founding member of the TRPN sub-family7, fulfills all the criteria for a bona fide mechanotransduction channel1,8, and is important for a variety of mechanosensation-related behaviors such as locomotion, touch and sound sensation across different species including C. elegans9, Drosophila8,10-11 and zebrafish12. NOMPC has 29 ARs, the largest number among TRP channels. They are implicated as tether to convey force from cytoskeleton to the channel, thus to mediate mechanosensation6,13-15. A key question is how the long AR domain is organized as a tether that can trigger channel gating. Here we present a de novo atomic structure of NOMPC determined by single particle electron cryo-microscopy (cryo-EM), and discuss how its architecture could provide a means to convey mechanical force to generating an electrical signal within a cell.
scite is a Brooklyn-based organization that helps researchers better discover and understand research articles through Smart Citations–citations that display the context of the citation and describe whether the article provides supporting or contrasting evidence. scite is used by students and researchers from around the world and is funded in part by the National Science Foundation and the National Institute on Drug Abuse of the National Institutes of Health.
