scholarly journals Intraflagellar transport drives flagellar surface motility

eLife ◽  
2013 ◽  
Vol 2 ◽  
Author(s):  
Sheng Min Shih ◽  
Benjamin D Engel ◽  
Fatih Kocabas ◽  
Thomas Bilyard ◽  
Arne Gennerich ◽  
...  
Keyword(s):  
Intraflagellar Transport ◽  
Gliding Motility ◽  
Solid Surfaces ◽  
Dependent Manner ◽  
Surface Adhesion ◽  
Membrane Glycoproteins ◽  
Flagellar Membrane ◽  
Surface Motility ◽  
Cilia And Flagella ◽  
Ciliary Signaling

The assembly and maintenance of all cilia and flagella require intraflagellar transport (IFT) along the axoneme. IFT has been implicated in sensory and motile ciliary functions, but the mechanisms of this relationship remain unclear. Here, we used Chlamydomonas flagellar surface motility (FSM) as a model to test whether IFT provides force for gliding of cells across solid surfaces. We show that IFT trains are coupled to flagellar membrane glycoproteins (FMGs) in a Ca2+-dependent manner. IFT trains transiently pause through surface adhesion of their FMG cargos, and dynein-1b motors pull the cell towards the distal tip of the axoneme. Each train is transported by at least four motors, with only one type of motor active at a time. Our results demonstrate the mechanism of Chlamydomonas gliding motility and suggest that IFT plays a major role in adhesion-induced ciliary signaling pathways.

scholarly journals The Chlamydomonas kinesin-like protein FLA10 is involved in motility associated with the flagellar membrane.

1995 ◽  
Vol 131 (6) ◽  
pp. 1517-1527 ◽  
Author(s):  
K G Kozminski ◽  
P L Beech ◽  
J L Rosenbaum
Keyword(s):  
Electron Microscopy ◽  
Light Microscopy ◽  
Intraflagellar Transport ◽  
Temperature Sensitive ◽  
Polystyrene Beads ◽  
Cell Biol ◽  
Flagellar Membrane ◽  
Surface Motility ◽  
Sensitive Allele ◽  
Bidirectional Movement

The Chlamydomonas FLA10 gene was shown to encode a flagellar kinesin-like protein (Walther, Z., M. Vashishtha, and J.L. Hall. 1994. J. Cell Biol. 126:175-188). By using a temperature-sensitive allele of FLA10, we have determined that the FLA10 protein is necessary for both the bidirectional movement of polystyrene beads on the flagellar membrane and intraflagellar transport (IFT), the bidirectional movement of granule-like particles beneath the flagellar membrane (Kozminski, K.G., K.A. Johnson, P. Forscher, and J.L. Rosenbaum. 1993. Proc. Natl. Acad. Sci. (USA). 90:5519-5523). In addition, we have correlated the presence and position of the IFT particles visualized by light microscopy with that of the electron dense complexes (rafts) observed beneath the flagellar membrane by electron microscopy. A role for FLA10 in submembranous or flagellar surface motility is also strongly supported by the immunolocalization of FLA10 to the region between the axonemal outer doublet microtubules and the flagellar membrane.

scholarly journals Flagella Ca2+ elevations regulate pausing of retrograde intraflagellar transport trains in adherent Chlamydomonas flagella

2020 ◽  
Author(s):  
Cecile Fort ◽  
Peter Collingridge ◽  
Colin Brownlee ◽  
Glen Wheeler
Keyword(s):  
Membrane Proteins ◽  
Green Alga ◽  
High Frequency ◽  
Intraflagellar Transport ◽  
Gliding Motility ◽  
Membrane Glycoproteins ◽  
Ciliary Membrane ◽  
Process Uncertainty ◽  
Temporal Properties ◽  
Transient Interactions

AbstractThe movement of ciliary membrane proteins is directed by transient interactions with intraflagellar transport (IFT) trains. The green alga Chlamydomonas has adapted this process for gliding motility, using IFT to move adhesive glycoproteins (FMG-1B) in the flagella membrane. Although Ca2+ signalling contributes directly to the gliding process, uncertainty remains over the mechanisms through which Ca2+ acts to influence the movement of IFT trains. Here we show that flagella Ca2+ elevations regulate IFT primarily by initiating the movement of paused retrograde IFT trains. Flagella Ca2+ elevations exhibit complex spatial and temporal properties, including high frequency repetitive Ca2+ elevations that prevent the accumulation of paused retrograde IFT trains. We show that flagella Ca2+ elevations disrupt the IFT-dependent movement of microspheres along the flagella membrane. The results suggest that flagella Ca2+ elevations directly disrupt the interaction between retrograde IFT particles and flagella membrane glycoproteins to modulate gliding motility and the adhesion of the flagellum to a surface.

Calcium influx regulates antibody-induced glycoprotein movements within the Chlamydomonas flagellar membrane

1990 ◽  
Vol 96 (1) ◽  
pp. 27-33 ◽  
Author(s):  
R.A. Bloodgood ◽  
N.L. Salomonsky
Keyword(s):  
Calcium Channel ◽  
Calcium Channel Blockers ◽  
Calcium Influx ◽  
Gliding Motility ◽  
Free Calcium ◽  
Membrane Glycoprotein ◽  
Channel Blockers ◽  
Membrane Glycoproteins ◽  
Whole Cell ◽  
Flagellar Membrane

The Chlamydomonas flagellar surface exhibits a number of dynamic membrane phenomena associated with whole-cell gliding locomotion and the early events in fertilization. Crosslinking of a specific population of flagellar surface-exposed glycoproteins with the lectin concanavalin A or an anti-carbohydrate mouse monoclonal antibody, designated FMG-1, results in a characteristic pattern of glycoprotein redistribution within the plane of the flagellar membrane. Recent evidence suggests that flagellar membrane glycoprotein movements are associated with both whole-cell gliding motility and the early events in mating. It is of interest to determine the transmembrane signaling pathway whereby crosslinking of the external domains of flagellar glycoproteins activates the intraflagellar machinery responsible for translocation of flagellar membrane glycoproteins. The redistribution of flagellar membrane glycoproteins requires micromolar levels of free calcium in the medium; lowering the free calcium concentration to 10(−7) M results in complete but reversible inhibition of redistribution. Redistribution is maximal in the presence of 20 microM free calcium in the medium. Redistribution is inhibited in the presence of 20 microM free calcium by the calmodulin antagonists trifluoperazine, W-7 and calmidazolium, the calcium channel blockers diltiazem, methoxyverapamil (D-600) and barium chloride, and the local anesthetics, lidocaine and procaine. The actions of all of these agents can be interpreted in terms of a requirement for calcium in the signaling mechanism associated with flagellar glycoprotein redistribution. In particular, the requirement for micromolar calcium in the external medium and the effects of specific calcium channel blockers suggest that flagellar membrane glycoprotein crosslinking may induce an increase in calcium influx, which may be the initial trigger for activating the flagellar machinery responsible for active movement of flagellar membrane glycoproteins.

scholarly journals The role of the dynein light intermediate chain in retrograde IFT and flagellar function in Chlamydomonas

2016 ◽  
Vol 27 (15) ◽  
pp. 2404-2422 ◽  
Author(s):  
Jaimee Reck ◽  
Alexandria M. Schauer ◽  
Kristyn VanderWaal Mills ◽  
Raqual Bower ◽  
Douglas Tritschler ◽  
...  
Keyword(s):  
Intraflagellar Transport ◽  
Small Subset ◽  
Dependent Manner ◽  
Microtubule Motor ◽  
Flagellar Assembly ◽  
Cilia And Flagella ◽  
The Stability ◽  
Mutant Phenotypes ◽  
Intermediate Chain

The assembly of cilia and flagella depends on the activity of two microtubule motor complexes, kinesin-2 and dynein-2/1b, but the specific functions of the different subunits are poorly defined. Here we analyze Chlamydomonas strains expressing different amounts of the dynein 1b light intermediate chain (D1bLIC). Disruption of D1bLIC alters the stability of the dynein 1b complex and reduces both the frequency and velocity of retrograde intraflagellar transport (IFT), but it does not eliminate retrograde IFT. Flagellar assembly, motility, gliding, and mating are altered in a dose-dependent manner. iTRAQ-based proteomics identifies a small subset of proteins that are significantly reduced or elevated in d1blic flagella. Transformation with D1bLIC-GFP rescues the mutant phenotypes, and D1bLIC-GFP assembles into the dynein 1b complex at wild-type levels. D1bLIC-GFP is transported with anterograde IFT particles to the flagellar tip, dissociates into smaller particles, and begins processive retrograde IFT in <2 s. These studies demonstrate the role of D1bLIC in facilitating the recycling of IFT subunits and other proteins, identify new components potentially involved in the regulation of IFT, flagellar assembly, and flagellar signaling, and provide insight into the role of D1bLIC and retrograde IFT in other organisms.

scholarly journals Ca2+ elevations disrupt interactions between intraflagellar transport and the flagella membrane in Chlamydomonas

2021 ◽  
Vol 134 (3) ◽  
pp. jcs253492
Author(s):  
Cecile Fort ◽  
Peter Collingridge ◽  
Colin Brownlee ◽  
Glen Wheeler
Keyword(s):  
Membrane Proteins ◽  
Green Alga ◽  
High Frequency ◽  
Traction Force ◽  
Intraflagellar Transport ◽  
Gliding Motility ◽  
Membrane Glycoproteins ◽  
Ciliary Membrane ◽  
Precise Control ◽  
Transient Interactions

ABSTRACTThe movement of ciliary membrane proteins is directed by transient interactions with intraflagellar transport (IFT) trains. The green alga Chlamydomonas has adapted this process for gliding motility, using retrograde IFT motors to move adhesive glycoproteins in the flagella membrane. Ca2+ signalling contributes directly to the gliding process, although uncertainty remains over the mechanism through which it acts. Here, we show that flagella Ca2+ elevations initiate the movement of paused retrograde IFT trains, which accumulate at the distal end of adherent flagella, but do not influence other IFT processes. On highly adherent surfaces, flagella exhibit high-frequency Ca2+ elevations that prevent the accumulation of paused retrograde IFT trains. Flagella Ca2+ elevations disrupt the IFT-dependent movement of microspheres along the flagella membrane, suggesting that Ca2+ acts by directly disrupting an interaction between retrograde IFT trains and flagella membrane glycoproteins. By regulating the extent to which glycoproteins on the flagella surface interact with IFT motor proteins on the axoneme, this signalling mechanism allows precise control of traction force and gliding motility in adherent flagella.

scholarly journals Pectin Induced Colony Expansion of Soil-Derived Flavobacterium Strains

2021 ◽  
Vol 12 ◽  
Author(s):  
Judith Kraut-Cohen ◽  
Orr H. Shapiro ◽  
Barak Dror ◽  
Eddie Cytryn
Keyword(s):  
Gliding Motility ◽  
Rapid Expansion ◽  
Solid Surfaces ◽  
Sugar Uptake ◽  
Dependent Manner ◽  
Plant Cell Walls ◽  
Plant Interactions ◽  
Related Proteins ◽  
Complex Organic ◽  
Plant Surfaces

The genus Flavobacterium is characterized by the capacity to metabolize complex organic compounds and a unique gliding motility mechanism. Flavobacteria are often abundant in root microbiomes of various plants, but the factors contributing to this high abundance are currently unknown. In this study, we evaluated the effect of various plant-associated poly- and mono-saccharides on colony expansion of two Flavobacterium strains. Both strains were able to spread on pectin and other polysaccharides such as microcrystalline cellulose. However, only pectin (but not pectin monomers), a component of plant cell walls, enhanced colony expansion on solid surfaces in a dose- and substrate-dependent manner. On pectin, flavobacteria exhibited bi-phasic motility, with an initial phase of rapid expansion, followed by growth within the colonized area. Proteomic and gene expression analyses revealed significant induction of carbohydrate metabolism related proteins when flavobacteria were grown on pectin, including selected SusC/D, TonB-dependent glycan transport operons. Our results show a positive correlation between colony expansion and the upregulation of proteins involved in sugar uptake, suggesting an unknown linkage between specific operons encoding for glycan uptake and metabolism and flavobacterial expansion. Furthermore, within the context of flavobacterial-plant interactions, they suggest that pectin may facilitate flavobacterial expansion on plant surfaces in addition to serving as an essential carbon source.

scholarly journals Gliding Motility of Mycoplasma mobile Can Occur by Repeated Binding to N-Acetylneuraminyllactose (Sialyllactose) Fixed on Solid Surfaces

2006 ◽  
Vol 188 (18) ◽  
pp. 6469-6475 ◽  
Author(s):  
Ryoichiro Nagai ◽  
Makoto Miyata
Keyword(s):  
Bovine Serum Albumin ◽  
Serum Albumin ◽  
Bovine Serum ◽  
Gliding Motility ◽  
Solid Surfaces ◽  
Dependent Manner ◽  
Animal Cells ◽  
Unknown Mechanism ◽  
Direct Binding ◽  
Lines Of Evidence

ABSTRACT Mycoplasma mobile relies on an unknown mechanism to glide across solid surfaces including glass, animal cells, and plastics. To identify the direct binding target, we examined the factors that affect the binding of Mycoplasma pneumoniae to solid surfaces and concluded that N-acetylneuraminyllactose (sialyllactose) attached to a protein can mediate glass binding on the basis of the following four lines of evidence: (i) glass binding was inhibited by N-acetylneuraminidase, (ii) glass binding was inhibited by N-acetylneuraminyllactose in a structure-dependent manner, (iii) binding occurred on glass pretreated with bovine serum albumin attached to N-acetylneuraminyllactose, and (iv) gliding speed depended on the density of N-acetylneuraminyllactose on glass.

scholarly journals A Dynein Light Chain Is Essential for the Retrograde Particle Movement of Intraflagellar Transport (IFT)

1998 ◽  
Vol 141 (4) ◽  
pp. 979-992 ◽  
Author(s):  
Gregory J. Pazour ◽  
Curtis G. Wilkerson ◽  
George B. Witman
Keyword(s):  
Light Chain ◽  
Intraflagellar Transport ◽  
Cytoplasmic Dynein ◽  
Dynein Light Chain ◽  
Particle Movement ◽  
Cell Biol ◽  
Flagellar Membrane ◽  
Radial Spokes ◽  
Surface Motility ◽  
Bidirectional Movement

Several enzymes, including cytoplasmic and flagellar outer arm dynein, share an Mr 8,000 light chain termed LC8. The function of this chain is unknown, but it is highly conserved between a wide variety of organisms. We have identified deletion alleles of the gene (fla14) encoding this protein in Chlamydomonas reinhardtii. These mutants have short, immotile flagella with deficiencies in radial spokes, in the inner and outer arms, and in the beak-like projections in the B tubule of the outer doublet microtubules. Most dramatically, the space between the doublet microtubules and the flagellar membrane contains an unusually high number of rafts, the particles translocated by intraflagellar transport (IFT) (Kozminski, K.G., P.L. Beech, and J.L. Rosenbaum. 1995. J. Cell Biol. 131:1517–1527). IFT is a rapid bidirectional movement of rafts under the flagellar membrane along axonemal microtubules. Anterograde IFT is dependent on a kinesin whereas the motor for retrograde IFT is unknown. Anterograde IFT is normal in the LC8 mutants but retrograde IFT is absent; this undoubtedly accounts for the accumulation of rafts in the flagellum. This is the first mutation shown to specifically affect retrograde IFT; the fact that LC8 loss affects retrograde IFT strongly suggests that cytoplasmic dynein is the motor that drives this process. Concomitant with the accumulation of rafts, LC8 mutants accumulate proteins that are components of the 15-16S IFT complexes (Cole, D.G., D.R. Deiner, A.L. Himelblau, P.L. Beech, J.C. Fuster, and J.L. Rosenbaum. 1998. J. Cell Biol. 141:993–1008), confirming that these complexes are subunits of the rafts. Polystyrene microbeads are still translocated on the surface of the flagella of LC8 mutants, indicating that the motor for flagellar surface motility is different than the motor for retrograde IFT.

scholarly journals Dynamics of the IFT machinery at the ciliary tip

eLife ◽  
2017 ◽  
Vol 6 ◽  
Author(s):  
Alexander Chien ◽  
Sheng Min Shih ◽  
Raqual Bower ◽  
Douglas Tritschler ◽  
Mary E Porter ◽  
...  
Keyword(s):  
Basal Body ◽  
Full Range ◽  
Intraflagellar Transport ◽  
Feedback Mechanism ◽  
Conventional Imaging ◽  
Range Of Movement ◽  
Anterograde Transport ◽  
Traffic Jam ◽  
Negative Feedback Mechanism ◽  
Cilia And Flagella

Intraflagellar transport (IFT) is essential for the elongation and maintenance of eukaryotic cilia and flagella. Due to the traffic jam of multiple trains at the ciliary tip, how IFT trains are remodeled in these turnaround zones cannot be determined by conventional imaging. Using PhotoGate, we visualized the full range of movement of single IFT trains and motors in Chlamydomonas flagella. Anterograde trains split apart and IFT complexes mix with each other at the tip to assemble retrograde trains. Dynein-1b is carried to the tip by kinesin-II as inactive cargo on anterograde trains. Unlike dynein-1b, kinesin-II detaches from IFT trains at the tip and diffuses in flagella. As the flagellum grows longer, diffusion delays return of kinesin-II to the basal body, depleting kinesin-II available for anterograde transport. Our results suggest that dissociation of kinesin-II from IFT trains serves as a negative feedback mechanism that facilitates flagellar length control in Chlamydomonas.

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