Mapping of the interaction sites between apoptosis linked gene ALG-2 and HEED.
The apoptosis linked gene (ALG-2) is a 22 kDa Ca2+-binding protein of the penta EF-hand motif family. ALG-2 was discovered in a "death trap" assay using T-cell receptor-mediated apoptosis. Depletion of ALG-2 using an anti-sense ALG-2 construct inhibits apoptosis that is induced by several stimuli, such as staurosporin, dexamethason, Fas, and glucocorticoid. The Ca2+-dependent function of ALG-2 is consistent with the observation that the cytoplasmic Ca2+ level is elevated in apoptotic cells. We found that ALG-2 interacted specifically with the carboxy-terminal region of HEED using a yeast two-hybrid assay system. The mutants of HEED were constructed by deleting five WD repeat motifs one by one from the C-terminus of the protein. These mutants of ALG-2 were made by combining the EF hand Ca2+ binding motifs in various ways. Mapping of the interaction sites, using each of the mutants, revealed that the interaction between HEED and a third EF-hand motif of ALG-2 was stronger than the other combination.
Lee
K H
KH
Bio-Med RRC and Division of Life Sciences, Pai Chai University, Taejon, Korea.
Kim
E
E
Lee
J S
JS
Lee
K S
KS
Kim
J W
JW
D051017:Apoptosis Regulatory Proteins; D001483:Base Sequence; D002135:Calcium-Binding Proteins / Q000235:genetics* / Q000378:metabolism; D005786:Gene Expression Regulation; D006367:HeLa Cells; D006801:Humans; D009154:Mutation; D063151:Polycomb Repressive Complex 2; D017434:Protein Structure, Tertiary; D012097:Repressor Proteins / Q000235:genetics* / Q000378:metabolism; D017384:Sequence Deletion; D014158:Transcription, Genetic; D020798:Two-Hybrid System Techniques
D051017:Apoptosis Regulatory Proteins; D002135:Calcium-Binding Proteins; C491488:EED protein, human; C097753:PDCD6 protein, human; C490815:Pdcd6 protein, mouse; D012097:Repressor Proteins; D063151:Polycomb Repressive Complex 2