English (United Kingdom)

https://curated-unify.zendy.io/wp-json/zendy-region/v1/featured_content/oa?rat=en

https://curated-unify.zendy.io/wp-json/zendy-region/v1/highlighted_journal/

Global: Zendy Plus annual

Presents the access of premium content as premium feature

Premium Content

Presents the keyphrase highlighting as premium feature

Keyphrase Highlighting

Presents the summarisation as premium feature

Summarisation

Insights

Presents the pdf analysis as premium feature

PDF Analysis

Presents the zaia usage as premium feature

ZAIA

Global: Zendy Tools annual

Global: Zendy Free Plan

Global: Zendy Tools monthly

Global: Zendy Plus monthly

Background  The building of a cilium or flagellum requires molecular motors and associated proteins that allow the relocation of proteins from the cell body to the distal end and the return of proteins to the cell body in a process termed intraflagellar transport (IFT). IFT trains are carried out by kinesin and back to the cell body by dynein.    Methods  We used whole genome sequencing to identify the causative mutations for two temperature-sensitive flagellar assembly mutants in  Chlamydomonas  and validated the changes using reversion analysis. We examined the effect of these mutations on the localization of IFT81, an IFT complex B protein, the cytoplasmic dynein heavy chain (DHC1b), and the dynein light intermediate chain (D1bLIC).    Results  The strains,  fla18  and  fla24 , have mutations in kinesin-2 and cytoplasmic dynein, respectively. The  fla18  mutation alters the same glutamic acid (E 24 G) mutated in the  fla10-14  allele (E 24 K). The  fla18  strain loses flagella at 32?C more rapidly than the E 24 K allele but less rapidly than the  fla10-1  allele. The  fla18  mutant loses its flagella by detachment rather than by shortening. The  fla24  mutation falls in cytoplasmic dynein and changes a completely conserved amino acid (L 3243 P) in an alpha helix in the AAA5 domain. The  fla24  mutant loses its flagella by shortening within 6 hours at 32?C. DHC1b protein is reduced by 18-fold and D1bLIC is reduced by 16-fold at 21?C compared to wild-type cells. We identified two pseudorevertants (L 3243 S and L 3243 R), which remain flagellated at 32?C. Although  fla24  cells assemble full-length flagella at 21?C, IFT81 protein localization is dramatically altered. Instead of localizing at the basal body and along the flagella, IFT81 is concentrated at the proximal end of the flagella. The pseudorevertants show wild-type IFT81 localization at 21?C, but proximal end localization of IFT81 at 32?C.    Conclusions  The change in the AAA5 domain of the cytoplasmic dynein in  fla24  may block the recycling of IFT trains after retrograde transport. It is clear that different alleles in the flagellar motors reveal different functions and roles. Multiple alleles will be important for understanding structure-function relationships.

New mutations in flagellar motors identified by whole genome sequencing in Chlamydomonas