Actin-depolymerizing factor, ADF/cofilin, is essentially required in assembly of Leishmania flagellum.
ADF/cofilins are ubiquitous actin dynamics-regulating proteins that have been mainly implicated in actin-based cell motility. Trypanosomatids, e.g. Leishmania and Trypanosoma, which mediate their motility through flagellum, also contain a putative ADF/cofilin homologue, but its role in flagellar motility remains largely unexplored. We have investigated the role of this protein in assembly and motility of the Leishmania flagellum after knocking out the ADF/cofilin gene by targeted gene replacement. The resultant mutants were completely immotile, short and stumpy, and had reduced flagellar length and severely impaired beat. In addition, the assembly of the paraflagellar rod was lost, vesicle-like structures were seen throughout the length of the flagellum and the state and distribution of actin were altered. However, episomal complementation of the gene restored normal morphology and flagellar function. These results for the first time indicate that the actin dynamics-regulating protein ADF/cofilin plays a critical role in assembly and motility of the eukaryotic flagellum.
Tammana
T V Satish
TV
Division of Molecular and Structural Biology, Central Drug Research Institute, Lucknow 226001, India.
Sahasrabuddhe
Amogh A
AA
Mitra
Kalyan
K
Bajpai
Virendra K
VK
Gupta
Chhitar M
CM
D051339:Actin Depolymerizing Factors / Q000235:genetics / Q000378:metabolism*; D000199:Actins / Q000235:genetics / Q000378:metabolism; D000818:Animals; D002465:Cell Movement; D003001:Cloning, Molecular; D016054:DNA, Protozoan / Q000235:genetics; D051345:Destrin / Q000235:genetics / Q000378:metabolism*; D005407:Flagella / Q000648:ultrastructure*; D017353:Gene Deletion; D055786:Gene Knockout Techniques; D006367:HeLa Cells; D006801:Humans; D007893:Leishmania donovani / Q000235:genetics* / Q000378:metabolism; D008855:Microscopy, Electron, Scanning; D046529:Microscopy, Electron, Transmission; D009154:Mutation; D015800:Protozoan Proteins / Q000235:genetics / Q000378:metabolism*; D011817:Rabbits; D011994:Recombinant Proteins / Q000235:genetics / Q000378:metabolism
D051339:Actin Depolymerizing Factors; D000199:Actins; D016054:DNA, Protozoan; D051345:Destrin; D015800:Protozoan Proteins; D011994:Recombinant Proteins