Figure 5.
Fig. 5. CBLC interacts with the WW domains of AIP4. A,
yeast two-hybrid analysis of CBLC/AIP4 interaction using several
CBLC and AIP4 construct shows that the proline-rich C-terminal
region (50 amino acids long) of CBLC and the WW domains of AIP4
are required for the interaction. Fusion between GAL4 DBD and
CBLC wild type, mutant of the TKB (RK), mutant of the RING
domain (CA), TKB, or C-terminal proline-rich region were tested
for two-hybrid interaction in AH109 yeast in combination with
fusion between AD of GAL4 with AIP4 C2 or with
the four WW domains of AIP4. GAL4 DBD fusion with lamin and GAL4
AD were used as controls. +, positive interaction; , negative
interaction; ND, not determined. B, pull-down experiment using a
GST-WW domains and a lysate of COS-1 cells expressing EGFP-CBLC
confirms that the four AIP4 WW domains bind to CBLC. Controls
show a binding of EGFP-CBLC to a GST-GRB2 and absence of binding
to GST.
C2 or with
the four WW domains of AIP4. GAL4 DBD fusion with lamin and GAL4
AD were used as controls. +, positive interaction; 
, negative
interaction; ND, not determined. B, pull-down experiment using a
GST-WW domains and a lysate of COS-1 cells expressing EGFP-CBLC
confirms that the four AIP4 WW domains bind to CBLC. Controls
show a binding of EGFP-CBLC to a GST-GRB2 and absence of binding
to GST.