Figure 3. The ubiquitin conjugation system is present in flagella, and its activity increases during flagellar shortening. (A) 62.5 μg flagella isolated from wild-type cells was used for in vitro ubiquitination with or without added ATP or HA∷ubiquitin (HA-Ub) and incubated for 5 or 30 min at 30°C. Flagellar proteins were analyzed on an immunoblot probed with an antibody to HA. Monomeric HA∷ubiquitin is ∼10 kD and formed dimers and trimers. The molecular mass of ubiquitinated proteins ranged from 27 to 200 kD. (B) The in vitro ubiquitination assay was performed on control, resorbing, and regenerating flagella and showed an increase in activity in resorbing flagella and a decrease in regenerating flagella. The reactions were incubated for 1 h at 22°C. The immunoblot was probed with HA antibody. Tubulin and RSP3 were used as loading controls. Molecular masses are given in kilodaltons.
Figure 3.

The ubiquitin conjugation system is present in flagella, and its activity increases during flagellar shortening. (A) 62.5 μg flagella isolated from wild-type cells was used for in vitro ubiquitination with or without added ATP or HA∷ubiquitin (HA-Ub) and incubated for 5 or 30 min at 30°C. Flagellar proteins were analyzed on an immunoblot probed with an antibody to HA. Monomeric HA∷ubiquitin is ∼10 kD and formed dimers and trimers. The molecular mass of ubiquitinated proteins ranged from 27 to 200 kD. (B) The in vitro ubiquitination assay was performed on control, resorbing, and regenerating flagella and showed an increase in activity in resorbing flagella and a decrease in regenerating flagella. The reactions were incubated for 1 h at 22°C. The immunoblot was probed with HA antibody. Tubulin and RSP3 were used as loading controls. Molecular masses are given in kilodaltons.

This Feature Is Available To Subscribers Only

or Create an Account

Close Modal
Close Modal