Mouse DAM1 regulates pro-apoptotic activity of BLK in mammary epithelial cells.
Mouse mammary gland cDNA library was screened to search for BLK-interacting protein using yeast two-hybrid system, and a mouse DNA amplified in mammary carcinoma 1 (mDAM1) was obtained. mDAM1 cDNA contained a full coding region of 678bp encoding 225 amino acids with the predicted molecular mass of 26kDa. Comparison of the mouse to human DAM1 revealed 90 and 100% identities at the nucleotide and protein levels, respectively. A single 1.5kb transcript for mDAM1 was expressed in NMuMG mouse mammary epithelial cells. Through in vitro protein binding assay, interaction between mDAM1 and BLK was also confirmed. NMuMG cells, stably transfected and expressing mDAM1, promoted cell death under serum starvation condition. Transient coexpression of mDAM1 and BLK showed increased cell death compared to BLK expression alone in NMuMG cells. These results indicate that mDAM1 promotes mammary epithelial cell death and pro-apoptotic function of BLK.
Lee
Sulra
S
School of Agricultural Biotechnology, Seoul National University, 441-744, Suwon, South Korea.
Ha
Seckho
S
Chung
Myungil
M
Kim
Youngmin
Y
Choi
Yunjaie
Y
D048868:Adaptor Proteins, Signal Transducing; D000595:Amino Acid Sequence; D000818:Animals; D017209:Apoptosis; D051017:Apoptosis Regulatory Proteins; D001483:Base Sequence; D015152:Blotting, Northern; D002352:Carrier Proteins / Q000235:genetics / Q000378:metabolism*; D018797:Cell Cycle Proteins / Q000502:physiology*; D002478:Cells, Cultured; D004847:Epithelial Cells / Q000378:metabolism*; D005260:Female; D015723:Gene Library; D006801:Humans; D066298:In Vitro Techniques; D008321:Mammary Glands, Animal / Q000378:metabolism*; D051379:Mice; D008869:Microtubule-Associated Proteins / Q000502:physiology*; D024101:Mitochondrial Proteins; D008969:Molecular Sequence Data; D010957:Plasmids; D011485:Protein Binding; D019253:Proto-Oncogene Proteins c-bcl-2 / Q000235:genetics / Q000378:metabolism*; D029701:Saccharomyces cerevisiae Proteins; D014162:Transfection; D020798:Two-Hybrid System Techniques
D048868:Adaptor Proteins, Signal Transducing; D051017:Apoptosis Regulatory Proteins; C111741:Bik protein, mouse; D002352:Carrier Proteins; D018797:Cell Cycle Proteins; C116003:DAM1 protein, S cerevisiae; D008869:Microtubule-Associated Proteins; D024101:Mitochondrial Proteins; D019253:Proto-Oncogene Proteins c-bcl-2; D029701:Saccharomyces cerevisiae Proteins