RESUMEN
Ciliary outer-arm dynein (OAD) consists of heavy chains (HCs), intermediate chains (ICs), and light chains (LCs), of which HCs are the motor proteins that produce force. Studies using the green alga Chlamydomonas have revealed that ICs and LCs form a complex (IC/LC tower) at the base of the OAD tail and play a crucial role in anchoring OAD to specific sites on the microtubule. In this study, we isolated a novel slow-swimming Chlamydomonas mutant deficient in the IC2 protein. This mutation, E279K, is in the third of the seven WD repeat domains. No apparent abnormality was observed in electron microscope observations of axonemes or in SDS-PAGE analyses of dynein subunits. To explore the reason for the lowered motility in this mutant, in vitro microtubule sliding experiments were performed, which revealed that the motor activity of the mutant OAD was lowered. In particular, a large difference was observed between wild type (WT) and the mutant in the microtubule sliding velocity in microtubule bundles formed with the addition of OAD: ~35.3 µm/sec (WT) and ~4.3 µm/sec (mutant). From this and other results, we propose that IC2 in an OAD interacts with the ß HC of the adjacent OAD, and that an OAD-OAD interaction is important for efficient beating of cilia and flagella.Key words: cilia, axoneme, dynein heavy chain, cooperativity.
Asunto(s)
Chlamydomonas , Dineínas , Dineínas/genética , Dineínas/metabolismo , Microtúbulos/metabolismo , Axonema/metabolismo , Cilios/metabolismo , Flagelos/metabolismo , Chlamydomonas/genética , Chlamydomonas/metabolismo , MutaciónRESUMEN
The single-cell green alga Chlamydomonas reinhardtii possesses two α-tubulin genes (tua1 and tua2) and two ß-tubulin genes (tub1 and tub2), with the two genes in each pair encoding identical amino acid sequences. Here, we screened an insertional library to establish eight disruptants with defective tua2, tub1, or tub2 expression. Most of the disruptants did not exhibit major defects in cell growth, flagellar length, or flagellar regeneration after amputation. Because few tubulin mutants of C. reinhardtii have been reported to date, we then used our disruptants, together with a tua1 disruptant obtained from the Chlamydomonas Library Project (CLiP), to isolate tubulin-mutants resistant to the anti-tubulin agents propyzamide (pronamide) or oryzalin. As a result of several trials, we obtained 8 strains bearing 7 different α-tubulin mutations and 12 strains bearing 7 different ß-tubulin mutations. One of the mutations is at a residue similar to that of a mutation site known to confer drug resistance in human cancer cells. Some strains had the same amino acid substitutions as those reported previously in C. reinhardtii; however, the mutants with single tubulin genes showed slightly stronger drug-resistance than the previous mutants that express the mutated tubulin in addition to the wild-type tubulin. Such increased drug-resistance may have facilitated sensitive detection of tubulin mutation. Single-tubulin-gene disruptants are thus an efficient background of generating tubulin mutants for the study of the structure-function relationship of tubulin.
Asunto(s)
Chlamydomonas reinhardtii , Genes de Plantas , Mutación , Proteínas de Plantas , Tubulina (Proteína) , Chlamydomonas reinhardtii/genética , Chlamydomonas reinhardtii/metabolismo , Proteínas de Plantas/genética , Proteínas de Plantas/metabolismo , Tubulina (Proteína)/genética , Tubulina (Proteína)/metabolismoRESUMEN
HSP40s are regarded as cochaperones, perpetually shuttling client polypeptides to HSP70s for refolding. However, many HSP40s that are central for disparate processes diverge from this paradigm. To elucidate the noncanonical mechanisms, we investigated HSP40 in the radial spoke (RS) complex in flagella. Disruption of the gene by the MRC1 transposon in Chlamydomonas resulted in jerky flagella. Traditional electron microscopy, cryo-electron tomography, and sub-tomogram analysis revealed RSs of various altered morphologies that, unexpectedly, differed between the two RS species. This indicates that HSP40 locks the RS into a functionally rigid conformation, facilitating its interactions with the adjacent central pair apparatus for transducing locally varied mechanical feedback, which permits rhythmic beating. Missing HSP40, like missing RSs, could be restored in a tip-to-base direction when HSP40 mutants fused with a HSP40 donor cell. However, without concomitant de novo RS assembly, the repair was exceedingly slow, suggesting HSP40/RS-coupled intraflagellar trafficking and assembly. Biochemical analysis and modeling uncovered spoke HSP40's cochaperone traits. On the basis of our data, we propose that HSP40 accompanies its client RS precursor when traveling to the flagellar tip. Upon arrival, both refold in concert to assemble into the mature configuration. HSP40's roles in chaperoning and structural maintenance shed new light on its versatility and flagellar biology.
Asunto(s)
Flagelos/metabolismo , Proteínas del Choque Térmico HSP40/metabolismo , Axonema/metabolismo , Axonema/ultraestructura , Proteínas Bacterianas/metabolismo , Chlamydomonas , Elementos Transponibles de ADN/genética , Tomografía con Microscopio Electrónico , Flagelos/ultraestructura , Modelos Moleculares , Mutagénesis Insercional/genética , Mutación/genética , Unión ProteicaRESUMEN
Chlamydomonas flagella display surface motility such that small polystyrene beads (microspheres) attached to the flagellar membrane move bidirectionally along the flagellum. This surface motility enables cells to glide on a solid substrate to which they are attached by the flagellar surface. Previous studies suggested that microsphere movement and gliding motility result from the movement of transmembrane glycoprotein(s) within the plane of the plasma membrane, driven by intraflagellar transport (IFT), which utilizes cytoplasmic dynein and kinesin-2. However, it is not well understood how a cell can continuously glide in one direction further than a single flagellar length. Here we show that, during microsphere translocation on the flagella of a non-motile mutant, pf18, some flagellar glycoproteins, including FMG-1B and FAP113, detach from the membrane and attach to the microspheres. We propose that such relocation of surface glycoproteins underlies the ability to glide over a long distance. Surface motility is likely common to cilia/flagella of various organisms, as a similar microsphere movement is observed in the apical ciliary tuft in sea urchin embryos.
Asunto(s)
Membrana Celular/fisiología , Chlamydomonas reinhardtii/fisiología , Flagelos/fisiología , Glicoproteínas/fisiología , Microesferas , Animales , Cilios/fisiología , Locomoción , Erizos de Mar/embriologíaRESUMEN
We determined how the ciliary motor I1 dynein is transported. A specialized adapter, IDA3, facilitates I1 dynein attachment to the ciliary transporter called intraflagellar transport (IFT). Loading of IDA3 and I1 dynein on IFT is regulated by ciliary length.
Asunto(s)
Axonema/metabolismo , Chlamydomonas/metabolismo , Cilios/metabolismo , Dineínas/metabolismo , Flagelos/metabolismo , Cinesinas/metabolismo , Modelos Biológicos , Mutación , Proteínas de Plantas/metabolismo , Biosíntesis de Proteínas , Transporte de ProteínasRESUMEN
The outer arm dynein (OAD) complex is the main propulsive force generator for ciliary/flagellar beating. In Chlamydomonas and Tetrahymena, the OAD complex comprises three heavy chains (α, ß, and γ HCs) and >10 smaller subunits. Dynein light chain-1 (LC1) is an essential component of OAD. It is known to associate with the Chlamydomonas γ head domain, but its precise localization within the γ head and regulatory mechanism of the OAD complex remain unclear. Here Ni-NTA-nanogold labeling electron microscopy localized LC1 to the stalk tip of the γ head. Single-particle analysis detected an additional structure, most likely corresponding to LC1, near the microtubule-binding domain (MTBD), located at the stalk tip. Pull-down assays confirmed that LC1 bound specifically to the γ MTBD region. Together with observations that LC1 decreased the affinity of the γ MTBD for microtubules, we present a new model in which LC1 regulates OAD activity by modulating γ MTBD's affinity for the doublet microtubule.
Asunto(s)
Dineínas Axonemales/metabolismo , Microtúbulos/metabolismo , Chlamydomonas/enzimología , Chlamydomonas/metabolismo , Cilios/enzimología , Cilios/metabolismo , Flagelos/enzimología , Flagelos/metabolismo , Microscopía Electrónica/métodos , Microtúbulos/enzimología , Modelos Moleculares , Unión Proteica , Estructura Terciaria de Proteína , Proteínas Protozoarias/metabolismo , Tetrahymena/enzimología , Tetrahymena/metabolismoRESUMEN
Ciliary length control is an incompletely understood process essential for normal ciliary function. The flagella of Chlamydomonas mutants lacking multiple axonemal dyneins are shorter than normal; previously it was shown that this shortness can be suppressed by the mutation suppressor of shortness 1 (ssh1) via an unknown mechanism. To elucidate this mechanism, we carried out genetic analysis of ssh1 and found that it is a new allele of TPG2 (hereafter tpg2-3), which encodes FAP234 functioning in tubulin polyglutamylation in the axoneme. Similar to the polyglutamylation-deficient mutants tpg1 and tpg2-1, tpg2-3 axonemal tubulin has a greatly reduced level of long polyglutamate side chains. We found that tpg1 and tpg2-1 mutations also promote flagellar elongation in short-flagella mutants, consistent with a polyglutamylation-dependent mechanism of suppression. Double mutants of tpg1 or tpg2-1 and fla10-1, a temperature-sensitive mutant of intraflagellar transport, underwent slower flagellar shortening than fla10-1 at restrictive temperatures, indicating that the rate of tubulin disassembly is decreased in the polyglutamylation-deficient flagella. Moreover, α-tubulin incorporation into the flagellar tips in temporary dikaryons was retarded in polyglutamylation-deficient flagella. These results show that polyglutamylation deficiency stabilizes axonemal microtubules, decelerating axonemal disassembly at the flagellar tip and shifting the axonemal assembly/disassembly balance toward assembly.
Asunto(s)
Chlamydomonas/citología , Chlamydomonas/metabolismo , Flagelos/metabolismo , Péptido Sintasas/metabolismo , Tubulina (Proteína)/metabolismo , Dineínas Axonemales/genética , Dineínas Axonemales/metabolismo , Axonema/metabolismo , Chlamydomonas/genética , Cilios/metabolismo , Mutación , Ácido Poliglutámico/metabolismoRESUMEN
Motile cilia and flagella rapidly propagate bending waves and produce water flow over the cell surface. Their function is important for the physiology and development of various organisms including humans. The movement is based on the sliding between outer doublet microtubules driven by axonemal dyneins, and is regulated by various axonemal components and environmental factors. For studies aiming to elucidate the mechanism of cilia/flagella movement and regulation, Chlamydomonas is an invaluable model organism that offers a variety of mutants. This chapter introduces standard methods for studying Chlamydomonas flagellar motility including analysis of swimming paths, measurements of swimming speed and beat frequency, motility reactivation in demembranated cells (cell models), and observation of microtubule sliding in disintegrating axonemes. Most methods may be easily applied to other organisms with slight modifications of the medium conditions.
Asunto(s)
Chlamydomonas reinhardtii/fisiología , Cilios/fisiología , Flagelos/fisiología , Hidrodinámica , Locomoción/fisiología , Dineínas Axonemales/metabolismo , Movimiento Celular/fisiología , Dineínas/metabolismo , Microtúbulos/metabolismoRESUMEN
Flagellar dyneins are essential microtubule motors in eukaryotes, as they drive the beating motions of cilia and flagella. Unlike myosin and kinesin motors, the track binding mechanism of dyneins and the regulation between the strong and weak binding states remain obscure. Here we report the solution structure of the microtubule-binding domain of flagellar dynein-c/DHC9 (dynein-c MTBD). The structure reveals a similar overall helix-rich fold to that of the MTBD of cytoplasmic dynein (cytoplasmic MTBD), but dynein-c MTBD has an additional flap, consisting of an antiparallel b sheet. The flap is positively charged and highly flexible. Despite the structural similarity to cytoplasmic MTBD, dynein-c MTBD shows only a small change in the microtubule- binding affinity depending on the registry change of coiled coil-sliding, whereby lacks the apparent strong binding state. The surface charge distribution of dynein-c MTBD also differs from that of cytoplasmic MTBD, which suggests a difference in the microtubule-binding mechanism.
Asunto(s)
Chlamydomonas reinhardtii/metabolismo , Dineínas/química , Microtúbulos/metabolismo , Proteínas de Plantas/química , Sitios de Unión , Dineínas/metabolismo , Simulación de Dinámica Molecular , Proteínas de Plantas/metabolismo , Estructura Secundaria de ProteínaRESUMEN
Existence of cellular structures with specific size raises a fundamental question in biology: How do cells measure length? One conceptual answer to this question is by a molecular ruler, but examples of such rulers in eukaryotes are lacking. In this work, we identified a molecular ruler in eukaryotic cilia and flagella. Using cryo-electron tomography, we found that FAP59 and FAP172 form a 96-nanometer (nm)-long complex in Chlamydomonas flagella and that the absence of the complex disrupted 96-nm repeats of axonemes. Furthermore, lengthening of the FAP59/172 complex by domain duplication resulted in extension of the repeats up to 128 nm, as well as duplication of specific axonemal components. Thus, the FAP59/172 complex is the molecular ruler that determines the 96-nm repeat length and arrangements of components in cilia and flagella.
Asunto(s)
Dineínas Axonemales/química , Chlamydomonas/fisiología , Flagelos/fisiología , Dineínas Axonemales/genética , Dineínas Axonemales/ultraestructura , Chlamydomonas/ultraestructura , Cilios/fisiología , Cilios/ultraestructura , Células Eucariotas/fisiología , Células Eucariotas/ultraestructura , Flagelos/ultraestructura , Conformación ProteicaRESUMEN
Cilia and flagella contain nine outer doublet microtubules and a pair of central microtubules. The central pair of microtubules (CP) is important for cilia/flagella beating, as clearly shown by primary ciliary dyskinesia resulting from the loss of the CP. The CP is thought to regulate axonemal dyneins through interaction with radial spokes (RSs). However, the nature of the CP-RS interaction is poorly understood. Here we examine the appearance of CPs in the axonemes of a Chlamydomonas mutant, bld12, which produces axonemes with 8 to 11 outer-doublets. Most of its 8-doublet axonemes lack CPs. However, in the double mutant of bld12 and pf14, a mutant lacking the RS, most 8-doublet axonemes contain the CP. Thus formation of the CP apparently depends on the internal space limited by the outer doublets and RSs. In 10- or 11-doublet axonemes, only 3-5 RSs are attached to the CP and the doublet arrangement is distorted most likely because the RSs attached to the CP pull the outer doublets toward the axonemal center. The CP orientation in the axonemes varies in double mutants formed between bld12 and mutants lacking particular CP projections. The mutant bld12 thus provides the first direct and visual information about the CP-RS interaction, as well as about the mechanism of CP formation.
Asunto(s)
Chlamydomonas/metabolismo , Microtúbulos/metabolismo , Proteínas de Plantas/metabolismo , Axonema/metabolismo , Axonema/ultraestructura , Sitios de Unión , Chlamydomonas/genética , Cilios/metabolismo , Flagelos/metabolismo , Microscopía Electrónica , Microtúbulos/química , Microtúbulos/genética , Mutación , Proteínas de Plantas/químicaRESUMEN
This review outlines the current knowledge of the functional diversity of axonemal dyneins, as revealed by studies with the model organism Chlamydomonas. Axonemal dyneins, which comprise outer and inner dynein arms, power cilia and flagella beating by producing sliding movements between adjacent outer-doublet microtubules. Outer- and inner-arm dyneins have traditionally been considered similar in structure and function. However, recent evidence suggests that they differ rather strikingly in subunit composition, axonemal arrangement, and molecular motor properties. We posit that these arms make up two largely independent motile systems; whereas outer-arm dynein can generate axonemal beating by itself under certain conditions, inner-arm dynein can generate beating only in cooperation with the central pair/radial spokes. This conclusion is supported by genome analyses of various organisms. Outer-arm dynein appears to be particularly important for nodal cilia of mammalian embryos that function for determination of left-right body asymmetry.
Asunto(s)
Dineínas Axonemales/metabolismo , Chlamydomonas/metabolismo , Regulación de la Expresión Génica/fisiología , Animales , Dineínas Axonemales/genética , Chlamydomonas/genética , Flagelos/metabolismo , Movimiento , MutaciónRESUMEN
Outer arm dynein (OAD) in cilia and flagella is bound to the outer doublet microtubules every 24 nm. Periodic binding of OADs at specific sites is important for efficient cilia/flagella beating; however, the molecular mechanism that specifies OAD arrangement remains elusive. Studies using the green alga Chlamydomonas reinhardtii have shown that the OAD-docking complex (ODA-DC), a heterotrimeric complex present at the OAD base, functions as the OAD docking site on the doublet. We find that the ODA-DC has an ellipsoidal shape â¼24 nm in length. In mutant axonemes that lack OAD but retain the ODA-DC, ODA-DC molecules are aligned in an end-to-end manner along the outer doublets. When flagella of a mutant lacking ODA-DCs are supplied with ODA-DCs upon gamete fusion, ODA-DC molecules first bind to the mutant axonemes in the proximal region, and the occupied region gradually extends toward the tip, followed by binding of OADs. This and other results indicate that a cooperative association of the ODA-DC underlies its function as the OAD-docking site and is the determinant of the 24-nm periodicity.
Asunto(s)
Axonema/metabolismo , Dineínas/metabolismo , Sustancias Macromoleculares/metabolismo , Microtúbulos/metabolismo , Modelos Biológicos , Western Blotting , Cromatografía en Gel , Electroforesis en Gel de Poliacrilamida , Electroporación , Técnica del Anticuerpo Fluorescente , Microscopía Electrónica , Microscopía Fluorescente , Unión Proteica , Colorantes de Rosanilina , UltracentrifugaciónRESUMEN
Cilia/flagella are assembled and maintained by the process of intraflagellar transport (IFT), a highly conserved mechanism involving more than 20 IFT proteins. However, the functions of individual IFT proteins are mostly unclear. To help address this issue, we focused on a putative IFT protein TTC26/DYF13. Using live imaging and biochemical approaches we show that TTC26/DYF13 is an IFT complex B protein in mammalian cells and Chlamydomonas reinhardtii. Knockdown of TTC26/DYF13 in zebrafish embryos or mutation of TTC26/DYF13 in C. reinhardtii, produced short cilia with abnormal motility. Surprisingly, IFT particle assembly and speed were normal in dyf13 mutant flagella, unlike in other IFT complex B mutants. Proteomic and biochemical analyses indicated a particular set of proteins involved in motility was specifically depleted in the dyf13 mutant. These results support the concept that different IFT proteins are responsible for different cargo subsets, providing a possible explanation for the complexity of the IFT machinery. DOI: http://dx.doi.org/10.7554/eLife.01566.001.
Asunto(s)
Proteínas Algáceas/metabolismo , Proteínas Portadoras/metabolismo , Movimiento Celular , Chlamydomonas reinhardtii/metabolismo , Cilios/metabolismo , Flagelos/metabolismo , Péptidos y Proteínas de Señalización Intracelular/metabolismo , Proteínas de Plantas/metabolismo , Proteínas de Pez Cebra/metabolismo , Proteínas Algáceas/genética , Animales , Proteínas Portadoras/genética , Línea Celular , Chlamydomonas reinhardtii/genética , Embrión no Mamífero/metabolismo , Regulación del Desarrollo de la Expresión Génica , Técnicas de Silenciamiento del Gen , Genotipo , Péptidos y Proteínas de Señalización Intracelular/genética , Ratones , Mutación , Fenotipo , Proteínas de Plantas/genética , Transporte de Proteínas , Transfección , Pez Cebra , Proteínas de Pez Cebra/genéticaRESUMEN
Tubulin undergoes various posttranslational modifications, including polyglutamylation, which is catalyzed by enzymes belonging to the tubulin tyrosine ligase-like protein (TTLL) family. A previously isolated Chlamydomonas reinhardtii mutant, tpg1, carries a mutation in a gene encoding a homologue of mammalian TTLL9 and displays lowered motility because of decreased polyglutamylation of axonemal tubulin. Here we identify a novel tpg1-like mutant, tpg2, which carries a mutation in the gene encoding FAP234, a flagella-associated protein of unknown function. Immunoprecipitation and sucrose density gradient centrifugation experiments show that FAP234 and TTLL9 form a complex. The mutant tpg1 retains FAP234 in the cell body and flagellar matrix but lacks it in the axoneme. In contrast, tpg2 lacks both TTLL9 and FAP234 in all fractions. In fla10, a temperature-sensitive mutant deficient in intraflagellar transport (IFT), both TTLL9 and FAP234 are lost from the flagellum at nonpermissive temperatures. These and other results suggest that FAP234 functions in stabilization and IFT-dependent transport of TTLL9. Both TTLL9 and FAP234 are conserved in most ciliated organisms. We propose that they constitute a polyglutamylation complex specialized for regulation of ciliary motility.
Asunto(s)
Axonema/enzimología , Chlamydomonas reinhardtii/enzimología , Péptido Sintasas/metabolismo , Proteínas de Plantas/metabolismo , Secuencia de Aminoácidos , Chlamydomonas reinhardtii/citología , Secuencia Conservada , Citoplasma/enzimología , Estabilidad de Enzimas , Flagelos/enzimología , Unión Proteica , Procesamiento Proteico-Postraduccional , Transporte de Proteínas , Homología de Secuencia de Aminoácido , Tubulina (Proteína)/metabolismoRESUMEN
Outer arm dynein (OAD) is bound to specific loci on outer-doublet-microtubules by interactions at two sites: via intermediate chain 1 (IC1) and the outer dynein arm docking complex (ODA-DC). Studies using Chlamydomonas mutants have suggested that the individual sites have rather weak affinities for microtubules, and therefore strong OAD attachment to microtubules is achieved by their cooperation. To test this idea, we examined interactions between IC1, IC2 (another intermediate chain) and ODA-DC using recombinant proteins. Recombinant IC1 and IC2 were found to form a 1:1 complex, and this complex associated with ODA-DC in vitro. Binding of IC1 to mutant axonemes revealed that there are specific binding sites for IC1. From these data, we propose a novel model of OAD-outer doublet association.
Asunto(s)
Axonema/química , Chlamydomonas reinhardtii/citología , Dineínas/química , Flagelos/metabolismo , Proteínas de Plantas/química , Animales , Sitios de Unión , Cromatografía de Afinidad , Dineínas/biosíntesis , Dineínas/aislamiento & purificación , Proteínas de Plantas/biosíntesis , Proteínas de Plantas/aislamiento & purificación , Unión Proteica , Mapeo de Interacción de Proteínas , Células Sf9 , SpodopteraRESUMEN
Axonemal dyneins must be precisely regulated and coordinated to produce ordered ciliary/flagellar motility, but how this is achieved is not understood. We analyzed two Chlamydomonas reinhardtii mutants, mia1 and mia2, which display slow swimming and low flagellar beat frequency. We found that the MIA1 and MIA2 genes encode conserved coiled-coil proteins, FAP100 and FAP73, respectively, which form the modifier of inner arms (MIA) complex in flagella. Cryo-electron tomography of mia mutant axonemes revealed that the MIA complex was located immediately distal to the intermediate/light chain complex of I1 dynein and structurally appeared to connect with the nexin-dynein regulatory complex. In axonemes from mutants that lack both the outer dynein arms and the MIA complex, I1 dynein failed to assemble, suggesting physical interactions between these three axonemal complexes and a role for the MIA complex in the stable assembly of I1 dynein. The MIA complex appears to regulate I1 dynein and possibly outer arm dyneins, which are both essential for normal motility.
Asunto(s)
Movimiento Celular , Chlamydomonas reinhardtii/citología , Cilios/metabolismo , Secuencia Conservada , Dineínas/metabolismo , Complejos Multiproteicos/metabolismo , Proteínas de Plantas/metabolismo , Secuencia de Aminoácidos , Axonema/metabolismo , Secuencia de Bases , Chlamydomonas reinhardtii/genética , Chlamydomonas reinhardtii/metabolismo , Chlamydomonas reinhardtii/ultraestructura , Cilios/ultraestructura , Dineínas/química , Genes de Plantas , Microtúbulos/metabolismo , Modelos Biológicos , Modelos Moleculares , Datos de Secuencia Molecular , Mutación/genética , Fenotipo , Proteínas de Plantas/química , Proteínas de Plantas/genética , Nexinas de Proteasas/metabolismo , Unión Proteica , Estabilidad Proteica , Transporte de Proteínas , Secuencias Repetitivas de AminoácidoRESUMEN
The tpg1 mutant of Chlamydomonas lacks the tubulin polyglutamylase TTLL9 and is deficient in flagellar tubulin polyglutamylation. It exhibits slow swimming, whereas the double mutant with oda2 (a slow-swimming mutant that lacks outer-arm dynein) is completely nonmotile. Thus, tubulin polyglutamylation must be important for the functioning of inner-arm dynein(s). In this study, we show that the tpg1 mutation only slightly affects the motility of mutants that lack dynein "e," one of the seven species of major inner-arm dyneins, whereas it greatly reduces the motility of mutants lacking other inner-arm dynein species. This suggests that dynein e is the main target of motility regulation by tubulin polyglutamylation. Furthermore, the motility of various mutants in the background of the tpg1 mutation raises the possibility that tubulin polyglutamylation also affects the dynein regulatory complex, a dynein e-associated key regulator of flagellar motility, which possibly constitutes the interdoublet (nexin) link. Tubulin polyglutamylation thus may play a central role in the regulation of ciliary and flagellar motility. © 2012 Wiley Periodicals, Inc.