Document Detail

A direct repeat of N-type Ca2+ channel alpha1B gene functions as a negative regulatory element in HeLa cells.
MedLine Citation:
PMID:  12018859     Owner:  NLM     Status:  MEDLINE    
The expression of the N-type voltage-gated calcium channel alpha1B gene is restricted to neurons by a 5'-upstream region (-3992 to -1788) that contains negative regulatory element(s) that are active in non-neuronal cells. A 39 bp DNA element, which is repeated nine times in a head-to-tail fashion, was found within the same region. To examine whether this direct repeat (DR) may function as a negatively acting cis-regulatory element, several fusion plasmids, DR-110alpha1BLUC (1X), DR-SV40LUC (IX, 2X), in which one or two copies of the DR fragment were subcloned upstream of the homologous and heterologous promoters, were transiently transfected into HeLa and NS20Y cells. The promoter activity of DR-110alpha1BLUC (1X) decreased to approximately 17% of the 110alph(a1B)LUC construct in HeLa cells. The expression of the DR-SV40LUC (1X) and DR-SV40LUC (2X) plasmids was also reduced to 50 to 23% of the levels that were observed in the pGL2-Promoter in the same cells. However, no repression of the DR constructs was observed in NS20Y cells. An electrophoretic mobility shift assay showed that two DR-specific complexes were detected in HeLa cells, but not in NS20Y cells. In addition, Southwestern blotting revealed the presence of approximately 33 and 43 kDa proteins in HeLa cells. Overall, these results suggest that a 39 bp DNA element might act as repressor in non-neuron cells through the specific interactions of the DNA-proteins.
So Yun Park; Yong Tae Park; Kyoon Eon Kim; Myung Chull Rhee; Hee Jung Cho; Dong Sun Kim
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Publication Detail:
Type:  Journal Article; Research Support, Non-U.S. Gov't    
Journal Detail:
Title:  Molecules and cells     Volume:  13     ISSN:  1016-8478     ISO Abbreviation:  Mol. Cells     Publication Date:  2002 Apr 
Date Detail:
Created Date:  2002-05-20     Completed Date:  2003-01-29     Revised Date:  2006-11-15    
Medline Journal Info:
Nlm Unique ID:  9610936     Medline TA:  Mol Cells     Country:  Korea (South)    
Other Details:
Languages:  eng     Pagination:  341-6     Citation Subset:  IM    
Department of Anatomy, School of Medicine, Kyungpook National University, Daegu, Korea.
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MeSH Terms
Calcium Channels, N-Type / genetics*
Genes, Reporter
Hela Cells
Neurons / cytology,  physiology
Regulatory Sequences, Nucleic Acid*
Repetitive Sequences, Nucleic Acid / genetics*
Reg. No./Substance:
0/Calcium Channels, N-Type

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