|
|
|
|
|
|
|
||||
| Home Help Feedback Subscriptions Archive Search Table of Contents | ||||
|
Fig. 4. The sumoylation motifs of LIN-1 are necessary for the interaction with
MEP-1. (A) Schematic of MEP-1 with zinc-finger motifs (black) and
glutamine-rich region (gray) (Belfiore et
al., 2002). (B) The interactions between LA:LIN-1 fusion proteins
and MEP-1(155-859) fused to the GAL4 activation domain (GAL4AD) were measured
qualitatively using the yeast two-hybrid system. A (+) indicates robust
activation of a LexA-dependent lacZ reporter gene. (C) The
interactions between wild-type LA:LIN-1(1-64) or the indicated mutant and
MEP-1 were measured quantitatively. Bars represent the average LexA-dependent
ß-galactosidase activity and lines indicate the standard deviation of at
least six independent yeast transformants. The signal with LA:LIN-1(1-64) was
set to 100 relative light units (RLU); the signals with mutant proteins are
proportional. (D) To monitor expression of LA:LIN-1 proteins, we analyzed
protein extracts from transformed yeast by western blotting using an anti-LA
antibody. Lanes 1-14 correspond to LA fusion proteins listed as 1-14 in C. (E)
The interaction of MEP-1 with the indicated LA:LIN-1 fusion protein was
measured quantitatively. Bars represent the average of six independent yeast
transformants and lines indicate the standard deviation. The signal with
LA:LIN-1(158-180) was set to 100 RLU and signals with mutant proteins are
proportional.
|
