A novel form of actin in Leishmania: molecular characterisation, subcellular localisation and association with subpellicular microtubules

Sahasrabuddhe, Amogh A.; Bajpai, V.K.; Gupta, C. M.

doi:10.1016/j.molbiopara.2003.11.008

Cited by 66 publications

(96 citation statements)

References 36 publications

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“…Furthermore, in more than 90% of dividing COF -/-cells, continuous actin cables were present between the two daughter cells, suggesting an impairment of actin dynamics in these cells. To confirm these findings, we prepared cytoskeletons of COF +/+ , COF -/-and COF -/-Comp cells using NP-40 (Sahasrabuddhe et al, 2004), and then analyzed them by western blotting and immunofluorescence microscopy (Fig. 8).…”

Section: Cof-null Cells Show Defects In Basal Body Separation and Clementioning

confidence: 84%

“…In higher organisms, ADF/cofilin is essential for depolymerization of the actin contractile ring that forms between two daughter cells during cell division (Abe et al, 1996). However, in the case of trypanosomatid parasites, cytokinesis is primarily a microtubule-mediated process, and despite the presence of actin and several actin-binding proteins (Sahasrabuddhe et al, 2004;Nayak et al, 2005;Kapoor et al, 2008;Tammana et al, 2008;Katta et al, 2009), their role in these parasites was largely unknown. Our recent studies (Tammana et al, 2008;Sahasrabuddhe et al, 2009) show that the actin-binding proteins Cof and CRN12 have essential roles in flagellar assembly and microtubule remodeling, respectively.…”

Section: Discussionmentioning

confidence: 99%

“…Leishmania actin and Cof antibodies were prepared following our published procedure (Sahasrabuddhe et al, 2004;Tammana et al, 2008) and anti--tubulin and anti--tubulin antibodies were procured from Sigma. The primary antibodies were used in combination with appropriate Alexa Fluor 488-conjugated goat anti-mouse secondary antibodies (Molecular Probes, 1:1000) and Cy3-conjugated goat anti-rabbit secondary antibodies (Sigma, 1:1000).…”

Section: Fluorescence Microscopymentioning

confidence: 99%

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ADF/cofilin-driven actin dynamics in early events ofLeishmaniacell division

Tammana

Sahasrabuddhe

Bajpai

et al. 2010

Journal of Cell Science

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ADF/cofilin is an actin-dynamics-regulating protein that is required for several actin-based cellular processes such as cell motility and cytokinesis. A homologue of this protein has recently been identified in the protozoan parasite Leishmania, which has been shown to be essentially required in flagellum assembly and cell motility. However, the role of this protein in cytokinesis remains largely unknown. We show here that deletion of the gene encoding ADF/cofilin in these organisms results in several aberrations in the process of cell division. These aberrations include delay in basal body and kinetoplast separation, cleavage furrow progression and flagellar pocket division. In addition to these changes, the intracellular trafficking and actin dynamics are also adversely affected. All these abnormalities are, however, reversed by episomal complementation. Together, these results indicate that actin dynamics regulates early events in Leishmania cell division.

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Section: Cof-null Cells Show Defects In Basal Body Separation and Clementioning

confidence: 84%

Section: Discussionmentioning

confidence: 99%

Section: Fluorescence Microscopymentioning

confidence: 99%

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ADF/cofilin-driven actin dynamics in early events ofLeishmaniacell division

Tammana

Sahasrabuddhe

Bajpai

et al. 2010

Journal of Cell Science

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“…These results suggested that Leishmania CRN12 might be an integral component of the basal body, as well as of some other undefined structural component in the flagellar pocket region. As actin has been shown to be present in this region (Sahasrabuddhe et al, 2004), we carefully examined colocalization of CRN12 with actin in the detergenttreated cells. Our analysis revealed only a limited co-localization of actin with CRN12 in the region outside the basal body.…”

Section: Resultsmentioning

confidence: 99%

“…Anti Leishmania recombinant CRN12 antibodies in mice and rabbits and anti Leishmania recombinant actin antibodies in rabbits were raised and purified as described earlier (Nayak et al, 2005;Sahasrabuddhe et al, 2004). Antibodies against α,β-tubulins (monoclonals) and anti-tyrosinated α-tubulin antibodies were procured from ICN and Chemicon (USA), respectively.…”

Section: Antibodiesmentioning

confidence: 99%

AncientLeishmaniacoronin (CRN12) is involved in microtubule remodeling during cytokinesis

Sahasrabuddhe

Nayak

Gupta

2009

Journal of Cell Science

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In general, coronins play an important role in actin-based processes, and are expressed in a variety of eukaryotic cells, including Leishmania. Here, we show that Leishmania coronin preferentially distributes to the distal tip during cytokinesis, and interacts with microtubules through a microtubule-based motor, kinesin K39. We further show that reduction in coronin levels by 40-50% in heterozygous coronin mutants results in generation of bipolar cells (25-30%), specifically in the log phase, owing to unregulated growth of the corset microtubules. Further analysis of bipolar cells revealed that the main cause of generation of bipolar cell morphology is the intrusion of the persistently growing corset microtubules into the other daughter cell corset from the opposite direction. This defect in cytokinesis, however, disappears upon episomal gene complementation. Additionally, our attempts to prepare homozygous mutants were unsuccessful, as only the aneuploid cells survive the selection process. These results indicate that coronin regulates microtubule remodeling during Leishmania cytokinesis and is essentially required for survival of these parasites in culture.

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Oligomerization of coronin: Implication on actin filament length in Leishmania

et al. 2015

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Coronin proteins bind with actin filaments and participate in regulation of actin-dependent processes. These proteins contain a coiled-coil domain at their C-terminus, which is responsible for their dimeric or trimeric forms. However, the functional significance of these oligomeric configurations in organizing the actin cytoskeleton is obscure. Here, we report that the Leishmania coronin exists in a higher oligomeric form through its coiled-coil domain, the truncation of which ablates the ability of Leishmania coronin to assist actin-filament formation. F-actin co-sedimentation assay using purified proteins shows that the coiled-coil domain does not interact with actin-filaments and its absence does not abrogate actin-coronin interaction. Furthermore, it was shown that unlike other coronins, Leishmania coronin interacts with actin-filaments through its unique region. These results provided important insights into the role of coronin oligomerization in modulating actin-network.

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A novel form of actin in Leishmania: molecular characterisation, subcellular localisation and association with subpellicular microtubules

Cited by 66 publications

References 36 publications

ADF/cofilin-driven actin dynamics in early events ofLeishmaniacell division

ADF/cofilin-driven actin dynamics in early events ofLeishmaniacell division

AncientLeishmaniacoronin (CRN12) is involved in microtubule remodeling during cytokinesis

Oligomerization of coronin: Implication on actin filament length in Leishmania

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