title: Characterization of ciliobrevin A mediated dynein ATPase inhibition on flagellar motility of Leishmania donovani.
abstract: Axonemal dyneins are members of AAA+ proteins involved in force generation and are responsible for flagellar motility in eukaryotes. In this study, we characterized the effects of ciliobrevin A (CbA), a dynein ATPase inhibitor, on flagella driven motility of the protozoan parasite Leishmania donovani. Using fast-capture video microscopy, we observed that CbA decreased flagellar beat frequency of swimming parasites in a concentration-dependent manner. Beat frequency of live and reactivated L. donovani decreased by approximately 89% and 41% respectively in the presence of 250μM CbA. This inhibition was lost when CbA was removed, suggesting its effects were reversible. CbA also altered wavelength and amplitude of the flagellum of live parasites. Waveform analysis of live and reactivated L. donovani revealed that CbA significantly affected flagellar waveform by inducing non-uniform bends with the flagellum beating away from the cell axis. These results suggest that CbA sensitive dynein ATPases possibly are responsible for power generation and waveform maintenance of the flagellum of L. donovani. This ability to inhibit axonemal dyneins also emphasizes the use of dynein inhibitors as valuable tools in studying functional roles of axonemal dyneins of flagellated eukaryotes.
authors.affiliation: Kusuma School of Biological Sciences, Indian Institute of Technology Delhi, Hauz Khas, New Delhi 110016, India.
authors.forename: G Srinivas
authors.initials: GS
authors.lastname: Reddy
authors.affiliation: Kusuma School of Biological Sciences, Indian Institute of Technology Delhi, Hauz Khas, New Delhi 110016, India.
authors.forename: Aakash Gautam
authors.initials: AG
authors.lastname: Mukhopadhyay
authors.affiliation: Kusuma School of Biological Sciences, Indian Institute of Technology Delhi, Hauz Khas, New Delhi 110016, India. Electronic address: csdey@bioschool.iitd.ac.in.
authors.forename: Chinmoy Sankar
authors.initials: CS
authors.lastname: Dey
mesh_terms: D004398:Dyneins / Q000037:antagonists & inhibitors*; D005407:Flagella / Q000502:physiology*; D007893:Leishmania donovani / Q000187:drug effects* / Q000502:physiology*; D008124:Locomotion / Q000187:drug effects*; D018715:Microscopy, Video; D052999:Quinazolinones / Q000378:metabolism*
chemical_list: D052999:Quinazolinones; C000598071:ciliobrevin A; D004398:Dyneins
keywords: Ciliobrevin; Dynein ATPases; Flagellar motility; Leishmania