SILKWORM 30K PROTEIN INHIBITS ECDYSONE-INDUCED APOPTOSIS BY BLOCKING THE BINDING OF ULTRASPIRACLE TO ECDYSONE RECEPTOR-B1 IN CULTURED Bm5 CELLS
- Authors
- Kim, Mi Young; Song, Hwa Young; Kim, Jin Hee; Kim, Bo Yong; Park, Sun Whan; Sung, Dong Kyung; Park, Hun Hee; Jeon, Soung Hoo; Chung, In Sik; Lee, Bong Hee
- Issue Date
- 11월-2012
- Publisher
- WILEY-BLACKWELL
- Keywords
- 30K protein; apoptosis; Bm5 cells; (ecdysone); silkworm
- Citation
- ARCHIVES OF INSECT BIOCHEMISTRY AND PHYSIOLOGY, v.81, no.3, pp.136 - 147
- Indexed
- SCIE
SCOPUS
- Journal Title
- ARCHIVES OF INSECT BIOCHEMISTRY AND PHYSIOLOGY
- Volume
- 81
- Number
- 3
- Start Page
- 136
- End Page
- 147
- URI
- https://scholar.korea.ac.kr/handle/2021.sw.korea/107125
- DOI
- 10.1002/arch.21050
- ISSN
- 0739-4462
- Abstract
- This study investigates the mechanism through which increased 30K protein inhibits ecdysone-induced apoptosis in the Bm5 silkworm ovarian cell line. Treatment of Bm5 cells with 20-hydroxyecdysone (20E) after transfection with the pIZT/V5-His control vector triggered apoptosis, but 20E treatment did not trigger apoptosis in Bm5 cells transfected with the pIZT/30K/V5-His vector. To confirm its inhibitory effect on apoptosis, 30K protein was first purified from Escherichia coli transformed with a 30K expression vector and used to generate specific antibodies in mice. Anti-30K antiserum was used to confirm synthesis of the 30K protein in pIZT/30K/V5-His-transfected Bm5 cells and to detect 30K protein binding to the ecdysone receptor-B1 (EcR-B1). Anti-30K antiserum was used to immunoprecipitate protein complexes containing 30K from Bm5 cells transfected with pIZT/30K/V5-His vector and treated with 20E. We observed that 30K proteins bound primarily to the EcR-B1 and not to ultraspiracle (USP). Reciprocal immunoprecipitation of EcR-B1-containing complexes from Bm5 cells transfected with control pIZT/V5-His vector and treated with 20E showed that EcR-B1 bound to USP in the absence of 30K but did not bind to USP in pIZT/30K/V5-His-transfected Bm5 cells. These results demonstrate that 30K proteins block USP binding to EcR-B1 through formation of a 30K/EcR-B1 complex, resulting in inhibition of 20E-induced Bm5 cell apoptosis.
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Collections - College of Life Sciences and Biotechnology > Division of Life Sciences > 1. Journal Articles
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