TABLE 2.
Yeast two-hybrid characterization of the ASAP1-Src interactiona
| LexA plasmid | VP16 plasmid
|
|
|---|---|---|
| VP16 | VP16-ASAP1-SID | |
| LexA | − | − |
| LexA-lamin | − | − |
| LexA-c-Src-SH3 | − | + |
| LexA-c-Src | − | + |
| LexA-c-SrcΔSH3 | − | − |
| LexA-c-SrcKD | − | + |
| LexA-n-Src | − | − |
| LexA-n-Src-SH3 | − | − |
a
Two-hybrid assays for protein-protein interactions were carried out by transformation of LexA and VP16 plasmids into the L40 yeast strain, which carries a lacZ reporter gene. β-Galactosidase filter assays on dual transformants were scored + (strong interaction) or − (no interaction). The LexA fusions tested include the following: lamin (negative control); c-Src-SH3 (wild-type c-Src SH3 domain); c-Src (wild-type full-length c-Src); c-SrcΔSH3 (c-Src with SH3 domain deletion); c-SrcKD (kinase dead c-Src K295R mutant); n-Src (full-length neuronal Src); n-Src-SH3 (SH3 domain of neuronal Src). VP16-ASAP1-SID contains amino acids 697 to 850 from ASAP1a.