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Silencing of a putative inner arm dynein heavy chain results in flagellar immotility in Trypanosoma brucei.

Abstract：

:The Trypanosoma brucei flagellum controls motility and is crucial for cell polarity and division. Unique features of trypanosome motility suggest that flagellar beat regulation in this organism is unusual and worthy of study. The flagellar axoneme, required for motility, has a structure that is highly conserved among eukaryotes. Of the several dyneins in the axonemal inner arm complex, dynein f is thought to control flagellar waveform shape. A T. brucei gene predicted to encode the dynein f alpha heavy chain, TbDNAH10, was silenced using RNA interference in procyclic T. brucei cells. This resulted in immotile flagella, showing no movement except for occasional slight twitches at the tips. Cell growth slowed dramatically and cells were found in large clusters. Microscopic analysis of silenced cultures showed many cells with detached flagella, sometimes entangled between multiple cells. DAPI staining showed an increased frequency of mis-positioned kinetoplasts and multinucleate cells, suggesting that these cells experience disruption at an early cell cycle stage, probably secondary to the motility defect. TEM images showed apparently normal axonemes and no discernable defects in inner arm structure. This study demonstrates the use of RNAi as an effective method to study very large genes such as dynein heavy chains (HCs), and the immotility phenotype of these dynein knockdowns suggests that an intact inner arm is necessary for flagellar beating in T. brucei. Since analogous mutants in Chlamydomonas reinhardtii retain motility, this phenotype likely reflects differences in requirements for motility and/or dynein assembly between the two organisms and these comparative studies will help elucidate the mechanisms of flagellar beat regulation.

journal_name

Mol Biochem Parasitol

journal_title

Molecular and biochemical parasitology

authors

Springer AL,Bruhn DF,Kinzel KW,Rosenthal NF,Zukas R,Klingbeil MM

doi

10.1016/j.molbiopara.2010.09.005

subject

Has Abstract

pub_date

2011-01-01 00:00:00

pages

68-75

issue

1

eissn

0166-6851

issn

1872-9428

pii

S0166-6851(10)00249-5

journal_volume

175

pub_type

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