TY - JOUR
T1 - Role of Sp1 in transcription of human ATP2A2 gene in keratinocytes
AU - Takagi, Atsushi
AU - Nishiyama, Chiharu
AU - Maeda, Keiko
AU - Tokura, Tomoko
AU - Kawada, Hiroshi
AU - Kanada, Shunsuke
AU - Niwa, Yusuke
AU - Nakano, Nobuhiro
AU - Mayuzumi, Nobuyasu
AU - Nishiyama, Makoto
AU - Ikeda, Shigaku
AU - Okumura, Ko
AU - Ogawa, Hideoki
N1 - Funding Information:
We thank the members of Atopy Research Center, Department of Dermatology, and Department of Immunology for their helpful discussions regarding this paper. We are grateful to Drs Miyuki Takagi, Tomonobu Ito, Naomi Shimokawa, William Ng, Tatsuo Fukai, Ms Mutsuko Hara, Ms Kanako Fukuayama, and Mr Hokuto Yokoyama for technical assistance, and Ms Michiyo Matsumoto for secretarial assistance. This work was supported by a Grant-in-Aid for Scientific Research (c) (to CN and SI) from the Ministry of Education, Culture, Sports, Science and Technology of Japan.
PY - 2008/1
Y1 - 2008/1
N2 - The ATP2A2 gene encodes Ca2+-dependent ATPase, the dysfunction of which causes Darier disease. In this study, we analyzed the promoter structure of the human ATP2A2 gene using primary normal human keratinocytes (NHK). Reporter assays showed that deletion of -550/-529, -488/-472, -390/-362, or -42/-21 resulted in a significant decrease in human ATP2A2 promoter activity. Electrophoretic mobility shift assay (EMSA) showed that Sp1 is a transcription factor that binds to the -550/-529 and -488/-472 regions of the promoter. Chromatin immunoprecipitation (ChIP) assay demonstrated that Sp1, but not Sp3, binds to the promoter region of the ATP2A2 gene in NHK cells in vivo. Knockdown of Sp1 expression by small interfering RNA resulted in a marked reduction in ATP2A2 promoter activity and ATP2A2 mRNA levels in NHK, suggesting that Sp1 positively transactivates the ATP2A2 promoter in NHK. This is early evidence demonstrating that Sp1 plays an important and positive role in ATP2A2 gene expression in NHK in vivo and in vitro.
AB - The ATP2A2 gene encodes Ca2+-dependent ATPase, the dysfunction of which causes Darier disease. In this study, we analyzed the promoter structure of the human ATP2A2 gene using primary normal human keratinocytes (NHK). Reporter assays showed that deletion of -550/-529, -488/-472, -390/-362, or -42/-21 resulted in a significant decrease in human ATP2A2 promoter activity. Electrophoretic mobility shift assay (EMSA) showed that Sp1 is a transcription factor that binds to the -550/-529 and -488/-472 regions of the promoter. Chromatin immunoprecipitation (ChIP) assay demonstrated that Sp1, but not Sp3, binds to the promoter region of the ATP2A2 gene in NHK cells in vivo. Knockdown of Sp1 expression by small interfering RNA resulted in a marked reduction in ATP2A2 promoter activity and ATP2A2 mRNA levels in NHK, suggesting that Sp1 positively transactivates the ATP2A2 promoter in NHK. This is early evidence demonstrating that Sp1 plays an important and positive role in ATP2A2 gene expression in NHK in vivo and in vitro.
UR - http://www.scopus.com/inward/record.url?scp=37049006932&partnerID=8YFLogxK
U2 - 10.1038/sj.jid.5700937
DO - 10.1038/sj.jid.5700937
M3 - Article
C2 - 17597815
AN - SCOPUS:37049006932
SN - 0022-202X
VL - 128
SP - 96
EP - 103
JO - Journal of Investigative Dermatology
JF - Journal of Investigative Dermatology
IS - 1
ER -