Table I.
Yeast two-hybrid assay of BCL2L12 and Beclin-1 interaction with BCL2 family proteins and GSK3β.
| pGAL4-DBD vector(pAS2-1) | pGAL4-AD vector (pACT2)
|
||||||||
|---|---|---|---|---|---|---|---|---|---|
| BCL-XL(1-209) | BCL2(1-211) | Beclin-1(1-450) | Bax(1-171) | BCL2L12(70-266) | BCL2L12(70-334) | BCL2L12(90-229) | BCL2L12(153-191) | BCL2L12(90-229), I209Aa | |
| BCL2L12(70-266) | B+ | B+ | – | B+ | ND | ND | ND | ND | ND |
| BCL2L12(70-266), L213A | – | – | ND | ND | ND | ND | ND | ND | ND |
| Bax(1-171) | B+ | B+ | ND | ND | B+ | B+ | B+ | ND | B+ |
| Bax(1-171), L63A | – | – | ND | ND | ND | ND | ND | ND | ND |
| Mcl-1(1-327) | ND | ND | – | – | – | – | ND | ND | ND |
| GSK3β | ND | ND | – | ND | B+ | B+ | B+ | B+ | ND |
a
BCL2L12 (90-229) L213A, L216A, L217A, E218A, E220A were all unable to affect the binding between BCL2L12 and Bax (data are not shown in the Table since their results are identical to that of the I209A mutant). BCL2L12, BCL2-like 12; BCL2, BCL2 apoptosis regulator; GSK3β, glycogen synthase kinase 3β; Bax, BCL2 associated X; Mcl-1, induced myeloid leukemia cell differentiation protein MCL-1; B+, interaction; –, no interaction; ND, not done, interaction is not tested in this assay. pGAL4-AD, pGAL4 transcription activation domain, pGAL4-DBD, factor's DNA-binding domain.