Slow ADP‐dependent acceleration of microtubule translocation produced by an axonemal dynein

Kikushima, Kenji; Yagi, Toshiki; Kamiya, Ritsu

doi:10.1016/s0014-5793(04)00278-9

Cited by 46 publications

(40 citation statements)

References 17 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…Without ADP preincubation, MTs did not show smooth unidirectional gliding but displayed irregular back-and-forth movements immediately after the introduction of MTs and ATP into the chamber. These observations suggest that OAD is activated by ADP through a slow process, similar to other kinds of dyneins studied previously (31,32). Thus, ADP regulation of in vitro dynein activity, first observed in inner arm dyneins (29), is a feature shared by Chlamydomonas OAD.…”

Section: Adp Incubation Of Oad Enhances and Stabilizes Mt Gliding-supporting

confidence: 79%

Systematic Comparison of in Vitro Motile Properties between Chlamydomonas Wild-type and Mutant Outer Arm Dyneins Each Lacking One of the Three Heavy Chains

Furuta

Yagi

Yanagisawa

et al. 2009

Journal of Biological Chemistry

Self Cite

View full text Add to dashboard Cite

Outer arm dynein (OAD) of cilia and flagella contains two or three distinct heavy chains, each having a motor function. To elucidate their functional difference, we compared the in vitro motile properties of Chlamydomonas wild-type OAD containing the ␣, ␤, and ␥ heavy chains and three kinds of mutant OADs, each lacking one of the three heavy chains. For systematic comparison, a method was developed to introduce a biotin tag into a subunit, LC2, which served as the specific anchoring site on an avidin-coated glass surface. Wild-type OAD displayed microtubule gliding in the presence of ATP and ADP, with a maximal velocity of 5.0 m/s, which is approximately 1 ⁄ 4 of the microtubule sliding velocity in the axoneme. The duty ratio was estimated to be as low as 0.08. The absence of the ␤ heavy chain lowered both the gliding velocity and ATPase activity, whereas the absence of the ␥ heavy chain increased both activities. Strikingly, the absence of the ␣ heavy chain lowered the gliding velocity but increased the ATPase activity. Thus, the three heavy chains are likely to play distinct roles and regulate each other to achieve coordinated force production.The rhythmic beating of eukaryotic cilia and flagella is produced by a regulated interaction between axonemal dyneins and outer doublet microtubules (MTs).2 The axonemal dyneins are classified into inner arm dyneins (IADs) and an outer arm dynein (OAD) according to their positions on the outer doublet. Each dynein is an ATPase complex composed of one to three heavy chains (HCs) and several smaller subunits. The HCs are composed of a head domain that produces a sliding force through an ATP-sensitive interaction with MTs and a tail domain that is stably fixed to the outer doublet A-tubule. In many organisms, OAD exists as a single large molecular complex, whereas the IADs exist as multiple, smaller complexes. Studies using Chlamydomonas reinhardtii mutants showed that IADs are important for the generation of a proper flagellar waveform, whereas OAD is important for the generation of high beat frequency (1, 2), suggesting that different dynein species have distinct functions. Functional diversity is also found among the multiple HCs in a single OAD complex; mutants that lack any one of the three OAD HCs (␣, ␤, and ␥), as well as mutants lacking the entire OAD, display reduced motility, but the degree of the motility defects varies depending on the missing HC (3-5). Thus, each HC apparently has a distinct role in OAD function. An important challenge is to elucidate how different HCs share their roles in the OAD complex to produce the overall movement.In accordance with in vivo observations, assays of in vitro MT gliding on dynein-coated surfaces have revealed motility differences among different outer arm HCs. In sea urchin OAD, which contains two HCs (␣ and ␤), a partial assembly that contains the ␤ HC and an intermediate chain is capable of translocating MTs, whereas the ␣ HC is not (6 -8). In Chlamydomonas OAD, the velocity of in vitro MT gliding induced by partial OAD as...

show abstract

Section: Adp Incubation Of Oad Enhances and Stabilizes Mt Gliding-supporting

confidence: 79%

Systematic Comparison of in Vitro Motile Properties between Chlamydomonas Wild-type and Mutant Outer Arm Dyneins Each Lacking One of the Three Heavy Chains

Furuta

Yagi

Yanagisawa

et al. 2009

Journal of Biological Chemistry

Self Cite

View full text Add to dashboard Cite

show abstract

“…It has been demonstrated that ADP is involved in the regulation of flagellar motility and dynein activity (Kinoshita et al, 1995;Omoto et al, 1996;Yagi, 2000;Kikushima et al, 2004). Our study indicated that the presence of a very low concentration of ADP is essential for continuous beating at very low ATP concentrations.…”

Section: Introductionsupporting

confidence: 50%

“…ADP has been shown to be a factor enabling the non-motile Chlamydomonas flagella missing a part of the central pair or radial spokes to beat (Omoto et al, 1996). ADP also increases the velocity of microtubule sliding by isolated inner arm dynein (Yagi, 2000;Shiroguchi and Toyoshima, 2001;Kikushima et al, 2004). The motile activity of cytoplasmic dynein has been shown to be regulated by the condition of nucleotide binding (Kon et al, 2004).…”

Section: Importance Of Adp In Flagellar Oscillationmentioning

confidence: 99%

Induction of Beating by Imposed Bending or Mechanical Pulse in Demembranated, Motionless Sea Urchin Sperm Flagella at Very Low ATP Concentrations

Ishikawa

Shingyoji

2007

Cell Struct. Funct.

View full text Add to dashboard Cite

ABSTRACT. A basic feature of the movement of eukaryotic flagella is oscillation. Although flagellar oscillation is thought to be regulated by a self-regulatory feedback system including the mechanical signal of bending itself, the mechanism regulating the dynein motile activity to produce oscillation is not well understood. To elucidate the mechanism, we developed a new experimental system which allowed us to analyze the conditions necessary for the induction of oscillation. When a mechanical signal of bending or a pulse was applied by micromanipulation to a demembranated motionless sea urchin sperm flagellar axoneme at very low ATP concentrations (1-3 µM), a localized pair of bends was induced. The bend formation was often followed by further responses including propagation of the distal bend of paired bends, growth and propagation of the paired bends, and cyclical beating. The beating was induced at 2.0 µM or higher concentrations of ATP, but appeared even at 1.5 µM ATP if a few µM of ADP was also present. When the proximal half of a flagellum was attached to a microneedle, beating could not be induced in the distal free region at 2 µM ATP. These results suggest that mechanical signal is involved in the mechanism regulating the motile activity of dynein to produce oscillation. Our results also showed that the presence of a small amount of ADP and the axial difference along the flagellum are factors essential for the induction of flagellar oscillation.

show abstract

“…To further examine the effects of the fluorescent ATP and ADP analogues on the dynein motile activity, we studied the role of ADP which has been shown to increase the initial velocity of microtubule translocation at 0.1 mM ATP by inner arm dynein of Chlamydomonas attached to the glass surface [Kikushima et al, 2004]. We found a similar increase of the velocity of microtubule translocation in 21S dynein: at 1 mM ATP the mean velocity gradually increased over a range of *10 min (Fig.…”

Section: Effects Of Stable Binding Of Adp On the Motile Activity Of Dmentioning

confidence: 78%

“…These inhibitory effects of higher concentrations of ATP can be released by the presence of ADP [Kinoshita et al, 1995;Omoto et al, 1996]. The role of ADP that enhances the dynein motile activity has also been shown in microtubule gliding assay experiments of purified monomeric dynein [Yagi, 2000;Shiroguchi and Toyoshima, 2001;Kikushima et al, 2004] and in experiments in which mechanical bending induced beating in motionless demembranated flagella at very low ATP conditions [Ishikawa and Shingyoji, 2007]. These findings concerning the roles of ATP and ADP seem to be related to the structural uniqueness of the dynein molecule with its multiple nucleotide-binding sites.…”

Section: Introductionmentioning

confidence: 79%

The roles of noncatalytic ATP binding and ADP binding in the regulation of dynein motile activity in flagella

Inoue

Shingyoji

2007

Cell Motil. Cytoskeleton

View full text Add to dashboard Cite

The regulation of dynein activity to produce microtubule sliding in flagella has not been well understood. To gain more insight into the roles of ATP and ADP in the regulation, we examined the effects of fluorescent ATP analogues and fluorescent ADP analogues on the ATPase activity and motile activity of dynein. 21S dynein purified from the outer arms of sea urchin sperm flagella hydrolyzed BODIPY(R) FL ATP (FL-ATP) at 78% of the rate for ATP hydrolysis. FL-ATP at 0.1-1 mM, however, induced neither microtubule translocation on a dynein-coated glass surface nor sliding disintegration of elastase-treated axonemes. Direct observation of single molecules of the fluorescent analogues showed that both the ATP and ADP analogues were stably bound to dynein over several minutes (dissociation rates = 0.0038-0.0082/s). When microtubule translocation on 21S dynein was induced by ATP, the initial increase of the mean velocity was accelerated by preincubation of the dynein with ADP. Similar increase was also induced by the preincubation with the ADP analogues. Even after preincubation with ADP, FL-ATP did not induce sliding disintegration of elastase-treated axonemes. After preincubation with a nonhydrolyzable ATP analogue, AMPPNP (adenosine 5'-(beta:gamma-imido)triphosphate), however, FL-ATP induced sliding disintegration in approximately 10% of the axonemes. These results indicate that both noncatalytic ATP binding and stable ADP binding, in addition to ATP hydrolysis, are involved in the regulation of the chemo-mechanical transduction in axonemal dynein.

show abstract

Slow ADP‐dependent acceleration of microtubule translocation produced by an axonemal dynein

Cited by 46 publications

References 17 publications

Systematic Comparison of in Vitro Motile Properties between Chlamydomonas Wild-type and Mutant Outer Arm Dyneins Each Lacking One of the Three Heavy Chains

Systematic Comparison of in Vitro Motile Properties between Chlamydomonas Wild-type and Mutant Outer Arm Dyneins Each Lacking One of the Three Heavy Chains

Induction of Beating by Imposed Bending or Mechanical Pulse in Demembranated, Motionless Sea Urchin Sperm Flagella at Very Low ATP Concentrations

The roles of noncatalytic ATP binding and ADP binding in the regulation of dynein motile activity in flagella

Contact Info

Product

Resources

About