Molecular cloning of a human cDNA encoding a novel protein, DAD1, whose defect causes apoptotic cell death in hamster BHK21 cells.
The tsBN7 cell line, one of the mutant lines temperature sensitive for growth which have been isolated from the BHK21 cell line, was found to die by apoptosis following a shift to the nonpermissive temperature. The induced apoptosis was inhibited by a protein synthesis inhibitor, cycloheximide, but not by the bcl-2-encoded protein. By DNA-mediated gene transfer, we cloned a cDNA that complements the tsBN7 mutation. It encodes a novel hydrophobic protein, designated DAD1, which is well conserved (100% identical amino acids between humans and hamsters). By comparing the base sequences of the parental BHK21 and tsBN7 DAD1 cDNAs, we found that the DAD1-encoding gene is mutated in tsBN7 cells. The DAD1 protein disappeared in tsBN7 cells following a shift to the nonpermissive temperature, suggesting that loss of the DAD1 protein triggers apoptosis.
Nakashima
T
T
Department of Molecular Biology, Graduate School of Medical Science, Kyushu University, Maidashi, Japan.
Sekiguchi
T
T
Kuraoka
A
A
Fukushima
K
K
Shibata
Y
Y
Komiyama
S
S
Nishimoto
T
T
D000595:Amino Acid Sequence; D000818:Animals; D017209:Apoptosis / Q000235:genetics / Q000502:physiology*; D051017:Apoptosis Regulatory Proteins; D001483:Base Sequence; D002460:Cell Line; D002461:Cell Line, Transformed; D003001:Cloning, Molecular; D017124:Conserved Sequence; D006224:Cricetinae; D003513:Cycloheximide / Q000494:pharmacology; D004247:DNA; D005816:Genetic Complementation Test; D006367:HeLa Cells; D006801:Humans; D008565:Membrane Proteins; D008969:Molecular Sequence Data; D011506:Proteins / Q000235:genetics* / Q000502:physiology*; D017386:Sequence Homology, Amino Acid; D013696:Temperature; D029867:Xenopus Proteins; D014982:Xenopus laevis
D051017:Apoptosis Regulatory Proteins; C489988:DAD1 protein, Xenopus; C083331:DAD1 protein, human; D008565:Membrane Proteins; D011506:Proteins; D029867:Xenopus Proteins; D004247:DNA; D003513:Cycloheximide