Choi, WS Yoon, SY Chang, II Choi, EJ Rhim, H Jin, BK Oh, TH Krajewski, S Reed, JC Oh, YJ 2024-01-21T14:11:11Z 2024-01-21T14:11:11Z 2021-09-05 2000-04 0022-3042 https://pubs.kist.re.kr/handle/201004/141491 To examine the correlation between the structure of Bcl-2 and its inhibitory function of c-Jun N-terminal kinase (JNK) and caspase activity, we established a dopaminergic neuronal cell line, MN9D overexpressing Bcl-2 (MN9D/Bcl-2) or its structural mutants. The mutants comprised a point mutation in the BH1 (G145A; MN9D/BH1) or BH2 (W188A; MN9D/BH2) domain and a deletion mutation in the C-terminal (MN9D/C22), BH3 (MN9D/BH3), or BH4 (MN9D/BH4) domain, As determined by the TUNEL (terminal deoxynucleotidyltransferase nick end-labeling) and MTT [3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide] reduction assay, apoptotic death of MN9D/Neo cells reached 80-90% within 24 h in response to 1 mu M staurosporine. Upon staurosporine treatment, JNK activity increased six- to sevenfold over the basal level within 2-4 h. Treatment of MN9D/Neo with both staurosporine and a caspase inhibitor, Z-VAD, attenuated cell death without suppressing JNK activation. Both staurosporine-induced cell death and JNK activation were attenuated in MN9D/ Bcl-2. As determined by cleavage of poly(ADP-ribose) polymerase into 85 kDa, Bcl-2 blocked caspase activity as well. When cells overexpressing one of the Bcl-2 mutants were treated with staurosporine, death was attenuated in MN9D/BH1, MN9D/BH2, and MN9D/C22 but not in MN9D/BH3 and MN9D/BH4. Similarly, both JNK and caspase activation were blocked in MN9D/BH1, MN9D/ BH2, and MN9D/C22, whereas they were not suppressed in MN9D/BH3 and MN9D/BH4. Taken together, our data indicate that there exists a close structural and functional correlation of Bcl-2 to JNK and caspase activity in staurosporine-induced dopaminergic neuronal cell death. English LIPPINCOTT WILLIAMS & WILKINS PROGRAMMED CELL-DEATH POLY(ADP-RIBOSE) POLYMERASE SYMPATHETIC NEURONS CYTOCHROME-C PROTEIN DOMAIN BAX ACTIVATION SURVIVAL MITOCHONDRIA Correlation between structure of Bcl-2 and its inhibitory function of JNK and caspase activity in dopaminergic neuronal apoptosis Article 10.1046/j.1471-4159.2000.0741621.x 1 JOURNAL OF NEUROCHEMISTRY, v.74, no.4, pp.1621 - 1626 JOURNAL OF NEUROCHEMISTRY 74 4 1621 1626 scie scopus 000085951400033 2-s2.0-17144441040 Biochemistry & Molecular Biology Neurosciences Biochemistry & Molecular Biology Neurosciences & Neurology Article PROGRAMMED CELL-DEATH POLY(ADP-RIBOSE) POLYMERASE SYMPATHETIC NEURONS CYTOCHROME-C PROTEIN DOMAIN BAX ACTIVATION SURVIVAL MITOCHONDRIA apoptosis Bcl-2 homology domain MN9D Parkinson&apos s disease