Functional characterization of two splice variants of rat bad and their interaction with Bcl-w in sympathetic neurons.
Neuronal cell death is in many cases regulated by competitive interactions between pro- and antiapoptotic proteins of the Bcl-2 family. In this study we have identified two splice variants of the rat proapoptotic molecule Bad, which differ in their carboxy-terminal regions. Both splice variants of Bad interacted with the antiapoptotic molecule Bcl-w as shown by yeast two-hybrid assay and by co-immunoprecipitation experiments from transfected cells. mRNA expression for the two variants of bad were detected in all neonatal and adult rat tissues tested. Overexpression of either of the two isoforms of Bad in nerve growth factor (NGF)-maintained sympathetic neurons by microinjection induced the cell death of these neurons, which was neutralized by co-expression of Bcl-w. Overexpression of Bcl-w in sympathetic neurons also counteracted death induced by NGF deprivation, which was not reduced by co-expression of either of the two Bad variants. The results suggest that Bcl-w, Bad-alpha, and Bad-beta may participate in the regulation of apoptosis in the sympathetic nervous system.
Hamnér
S
S
Department of Neuroscience and Neurobiology, Uppsala University, Uppsala, 751 23, Sweden.
Arumäe
U
U
Li-Ying
Y
Y
Sun
Y F
YF
Saarma
M
M
Lindholm
D
D
D017398:Alternative Splicing / Q000235:genetics*; D000595:Amino Acid Sequence; D000818:Animals; D051017:Apoptosis Regulatory Proteins; D002352:Carrier Proteins / Q000008:administration & dosage / Q000235:genetics* / Q000378:metabolism*; D016923:Cell Death / Q000235:genetics; D002470:Cell Survival / Q000187:drug effects / Q000235:genetics; D002478:Cells, Cultured; D003001:Cloning, Molecular; D051379:Mice; D008845:Microinjections; D008969:Molecular Sequence Data; D020932:Nerve Growth Factor / Q000494:pharmacology; D009474:Neurons / Q000166:cytology / Q000187:drug effects / Q000378:metabolism*; D011233:Precipitin Tests; D020033:Protein Isoforms / Q000008:administration & dosage / Q000235:genetics / Q000378:metabolism; D011506:Proteins / Q000378:metabolism*; D019253:Proto-Oncogene Proteins c-bcl-2; D012333:RNA, Messenger / Q000096:biosynthesis; D051381:Rats; D013564:Sympathetic Nervous System / Q000166:cytology / Q000187:drug effects / Q000378:metabolism; D014162:Transfection; D020798:Two-Hybrid System Techniques; D051025:bcl-Associated Death Protein
D051017:Apoptosis Regulatory Proteins; C494820:Bad protein, mouse; C494821:Bad protein, rat; C101155:Bcl2l2 protein, mouse; C491072:Bcl2l2 protein, rat; D002352:Carrier Proteins; D020033:Protein Isoforms; D011506:Proteins; D019253:Proto-Oncogene Proteins c-bcl-2; D012333:RNA, Messenger; D051025:bcl-Associated Death Protein; D020932:Nerve Growth Factor