PKA-Mediated Regulation of B/K Gene Transcription in PC12 Cells.
- Author:
Mi Hyun CHOI
1
;
Ho Shik KIM
;
Sung Ho CHOI
;
Mi Young KIM
;
Yoon Seong JANG
;
Young Min JANG
;
Jeong Hwa LEE
;
Seong Whan JEONG
;
In Kyung KIM
;
Oh Joo KWON
Author Information
1. Department of Biochemistry, The Catholic University of Korea College of Medicine, Seoul 137-701, Korea. ojkwon@catholic.ac.kr
- Publication Type:Original Article
- Keywords:
B/K protein;
Cyclic AMP;
Protein kinase A;
PC12 cells;
Forskolin;
cAMP response element
- MeSH:
Animals;
Colforsin;
Cyclic AMP;
Cyclic AMP-Dependent Protein Kinases;
DNA;
Electrophoretic Mobility Shift Assay;
PC12 Cells*;
Promoter Regions, Genetic;
Receptors, Purinergic P1;
RNA, Messenger
- From:The Korean Journal of Physiology and Pharmacology
2005;9(6):333-339
- CountryRepublic of Korea
- Language:English
-
Abstract:
B/K protein is a novel protein containing double C2-like domains. We examined the specific signaling pathway that regulates the transcription of B/K in PC12 cells. When the cells were treated with forskolin (50microM), B/K mRNA and protein levels were time-dependently decreased, reaching the lowest level at 3 or 4 hr, and thereafter returning to the control level. Chemicals such as dibutyryl-cAMP, cell- permeable cyclic AMP (cAMP) analogue and CGS21680, adenosine receptor A2A agonist, also repressed the B/K transcription. However, 1, 9-dideoxyforskolin did not show inhibitory effect on B/K transcription, suggesting direct involvement of cAMP in the forskolin-induced inhibition of B/K transcription. Effect of forskolin, dibutyryl cAMP and CGS21680 was significantly reduced in PKA-deficient PC12 cell line (PC12-123.7). One cAMP-response element (CRE) -like sequence (B/K CLS) was found in the promoter region of B/K DNA, and electrophoretic mobility shift assay indicated its binding to CREM and CREB. Forskolin significantly suppressed the promoter activity in CHO-K1 cells transfected with the constructs containing B/K CLS, but not with the construct in which B/K CLS was mutated (AC: TG). Taken together, we suggest that the transcription of B/K gene in PC12 cells may be regulated by PKA-dependent mechanism.
