Anomalies in the motion dynamics of long-flagella mutants of Chlamydomonas reinhardtii

Khona, Dolly; Rao, Venkatramanan G.; Motiwalla, Mustafa J.; Varma, P. C. Sreekrishna; Kashyap, Anisha R.; Das, Koyel; Shirolikar, Seema; Borde, Lalit; Dharmadhikari, J. A.; Dharmadhikari, A. K.; Mukhopadhyay, Susanta; Mathur, D.; D’Souza, Jacinta S.

doi:10.1007/s10867-012-9282-8

Cited by 23 publications

(20 citation statements)

References 39 publications

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“…6A). Those results are compatible with the work of Khona, et al, on long-cilia mutants (44). In the prior study four lf mutants were analyzed and displayed a reduced beat frequency (by 8-16%) as well as a reduced swimming velocity (by 26-57%) for cilia length >18 µm.…”

Section: Limitationssupporting

confidence: 90%

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How does cilium length affect beating?

Bottier

Thomas

Dutcher

et al. 2018

Preprint

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The effects of cilium length on the dynamics of cilia motion were investigated by high-speed video microscopy of uniciliate mutants of the swimming alga, Chlamydomonas reinhardtii. Cells with short cilia were obtained by deciliating cells via pH shock and allowing cilia to reassemble for limited times.The frequency of cilia beating was estimated from motion of the cell body and of the cilium. Key features of the ciliary waveform were quantified from polynomial curves fitted to the cilium in each image frame. Most notably, periodic beating did not emerge until the cilium reached a critical length between 2-4 µm. Surprisingly, in cells that exhibited periodic beating, the frequency of beating was similar for all lengths with only a slight decrease in frequency as length increased from 4 µm to the normal length of 10-12 µm. The waveform average curvature (rad/µm) was also conserved as the cilium grew. The mechanical metrics of ciliary propulsion: force, torque, and power all increased in proportion to length. Mechanical efficiency of beating appeared to be maximal at the normal wild-type length of 10-12 μm. These quantitative features of ciliary behavior illuminate the biophysics of cilia motion and, in future studies, may help distinguish competing hypotheses of the underlying mechanism of oscillation.

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Section: Limitationssupporting

confidence: 90%

“…Long-cilia mutant. We did not study cilia behavior in extremely long cilia such as those of lf mutants (44,67,68). In the current study, using wild-type cells we observed that rotation rate and forces applied by the cilium increased with length.…”

Section: Limitationsmentioning

confidence: 99%

How does cilium length affect beating?

Bottier

Thomas

Dutcher

et al. 2018

Preprint

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“…The wild type (cc124) and mutant strains deficient in the RS ( pf14) and CP ( pf14 , pf15 , pf16 , pf18 , pf19, and pf20) and cc620, cc621 were procured from Chlamydomonas Resource Center (University of Minnesota). All cells were grown in liquid TAP medium with continuous light under shaking conditions at 25 °C [ 37 ].…”

Section: Methodsmentioning

confidence: 99%

Myc-binding protein orthologue interacts with AKAP240 in the central pair apparatus of the Chlamydomonas flagella

et al. 2016

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BackgroundFlagella and cilia are fine thread-like organelles protruding from cells that harbour them. The typical ‘9 + 2’ cilia confer motility on these cells. Although the mechanistic details of motility remain elusive, the dynein-driven motility is regulated by various kinases and phosphatases. A-kinase anchoring proteins (AKAPs) are scaffolds that bind to a variety of such proteins. Usually, they are known to possess a dedicated domain that in vitro interacts with the regulatory subunits (RI and RII) present in the cAMP-dependent protein kinase (PKA) holoenzyme. These subunits conventionally harbour contiguous stretches of a.a. residues that reveal the presence of the Dimerization Docking (D/D) domain, Catalytic interface domain and cAMP-Binding domain. The Chlamydomonas reinhardtii flagella harbour two AKAPs; viz., the radial spoke AKAP97 or RSP3 and the central pair AKAP240. Both these were identified on the basis of their RII-binding property. Interestingly, AKAP97 binds in vivo to two RII-like proteins (RSP7 and RSP11) that contain only the D/D domain.ResultsWe found a Chlamydomonas Flagellar Associated Protein (FAP174) orthologous to MYCBP-1, a protein that binds to organellar AKAPs and Myc onco-protein. An in silico analysis shows that the N-terminus of FAP174 is similar to those RII domain-containing proteins that have binding affinities to AKAPs. Binding of FAP174 was tested with the AKAP97/RSP3 using in vitro pull down assays; however, this binding was rather poor with AKAP97/RSP3. Antibodies were generated against FAP174 and the cellular localization was studied using Western blotting and immunoflourescence in wild type and various flagella mutants. We show that FAP174 localises to the central pair of the axoneme. Using overlay assays we show that FAP174 binds AKAP240 previously identified in the C2 portion of the central pair apparatus.ConclusionIt appears that the flagella of Chlamydomonas reinhardtii contain proteins that bind to AKAPs and except for the D/D domain, lack the conventional a.a. stretches of PKA regulatory subunits (RSP7 and RSP11). We add FAP174 to this growing list.Electronic supplementary materialThe online version of this article (doi:10.1186/s12860-016-0103-y) contains supplementary material, which is available to authorized users.

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“…Cells were grown and maintained by periodic transfers in Tris Acetate Phosphate (TAP; pH 7) medium as mentioned by Khona et al. . For all experiments, ~1 × 10 8 cells · mL −1 were exposed to different concentrations of menadione (5, 10, 50, 100, and 200 μM).…”

Section: Methodsmentioning

confidence: 99%

Menadione‐induced caspase‐dependent programmed cell death in the green chlorophyte Chlamydomonas reinhardtii

Vavilala

Sinha

D’Souza

2014

Journal of Phycology

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Menadione, a quinone that undergoes redox cycles leading to the formation of superoxide radicals, induces programmed cell death (PCD) in animals and plants. In this study, we investigated whether the unicellular green alga Chlamydomonas reinhardtii P.A.Dangeard is capable of executing PCD upon exposure to menadione stress. We report here, the morphological, molecular, and biochemical changes after menadione exposure of C. reinhardtii cells. The effect of menadione on cell death has been shown to be dose-dependent; 5-100 μM menadione causes 20%-46% cell death, respectively. It appears that growth is inhibited with the concomitant degradation of the photosynthetic pigments and by a decrease in the photosynthetic capacity. Being an oxidative stress, we found an H2 O2 burst within 15 min of menadione exposure, followed by an increase in antioxidant enzyme (superoxide dismutase [SOD], catalase [CAT], and ascorbate peroxidase [APX]) activities. In parallel, RT-PCR was performed for transcript analyses of Mn-SOD, CAT, and APX. Our results clearly revealed that expression of these genes were up-regulated upon menadione exposure. Furthermore, classical hallmarks of PCD such as alteration of mitochondrial membrane potential, significant increase in caspase-3-like DEVDase activity, cleavage of poly (ADP) ribose polymerase (PARP)-1-like enzyme, and DNA fragmentation as detected by terminal deoxynucleotidyl transferase-mediated dUTP nick end-labeling (TUNEL) assay and oligosomal DNA fragmentation were observed. Moreover, antibodies against a mammalian active caspase-3 shared epitopes with a caspase-3-like protein of ~17 kDa; its pattern of expression and activity correlated with the onset of cell death. To the best of our knowledge, this is the first report on menadione-induced PCD through a mitochondrian-caspase protease pathway in an algal species.

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Anomalies in the motion dynamics of long-flagella mutants of Chlamydomonas reinhardtii

Cited by 23 publications

References 39 publications

How does cilium length affect beating?

How does cilium length affect beating?

Myc-binding protein orthologue interacts with AKAP240 in the central pair apparatus of the Chlamydomonas flagella

Menadione‐induced caspase‐dependent programmed cell death in the green chlorophyte Chlamydomonas reinhardtii

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