TY - JOUR
T1 - Genomic organization, chromosomal localization, and promoter of human gene for FK506-binding protein 12.6
AU - Nakazawa, Tetsuya
AU - Takasawa, Shin
AU - Noguchi, Naoya
AU - Nata, Koji
AU - Tohgo, Akira
AU - Mori, Mitsuko
AU - Nakagawara, Kan Ichi
AU - Akiyama, Takako
AU - Ikeda, Takayuki
AU - Yamauchi, Akiyo
AU - Takahashi, Iwao
AU - Yoshikawa, Takeo
AU - Okamoto, Hiroshi
N1 - Funding Information:
We are grateful to Mr. Yuya Shichinohe for technical assistance and Mr. Brent Bell for critical reading of the manuscript. This work was supported in part by Grants-in-Aid for Scientific Research from the Ministry of Education, Culture, Sports, Science and Technology, Japan and is in partial fulfillment by T. N. of the degree of Doctor of Medical Science at Tohoku University. T. A. is the recipient of a fellowship from the Japan Society for Promotion of Science.
PY - 2005/10/24
Y1 - 2005/10/24
N2 - Cyclic ADP-ribose (cADPR) induces the release of Ca2+ from microsomes of pancreatic islets for insulin secretion. It has been demonstrated that cADPR binds to FK506-binding protein 12.6 (FKBP 12.6) on rat islet ryanodine receptor and that the binding of cADPR to FKBP12.6 frees the ryanodine receptor from FKBP12.6, causing it to release Ca2+ [Noguchi, N., Takasawa, S., Nata, K., Tohgo, A., Kato, I., Ikehata, F., Yonekura, H., Okamoto, H., 1997. Cyclic ADP-ribose binds to FK506-binding protein to release Ca 2+ from islet microsomes. J. Biol. Chem. 272, 3133-3136.]. In this study, we cloned, characterized the structural organization of the human FKBP12.6, which is highly homologous to human FKBP12, and analyzed the promoters for FKBP12.6 and FKBP12. Human FKBP12.6 gene spanned about 16 kb in length and consisted of four exons and three introns. The positions of exon-intron junction of the FKBP12.6 gene were perfectly matched with those of FKBP12 gene except that FKBP12 has an additional exon, exon V, to code exclusively for 3′-UTR. Fluorescence in situ hybridization revealed that the FKBP12.6 gene was located on chromosome 2 p21-23, which is different from the locus (chromosome 20 p13) of the FKBP12 gene. Reporter gene analyses revealed that the regions of - 58 ∼ - 24 of FKBP12.6 and - 106 ∼ - 79 of FKBP12 are important for promoter activities. The promoters contain a consensus transcription factor binding sequence for Sp family in FKBP12.6 and Ets-1 in FKBP12. Electrophoretic mobility shift assays showed that nuclear proteins bind to the promoters. The DNA/protein complex on FKBP12.6 promoter was competed out by Sp1 consensus probe and the complex was supershifted by anti-Sp3 antibodies. On the other hand, the DNA/protein complex on FKBP12 promoter was competed out by Ets-1 consensus probe but not by its mutant probe, indicating that Sp3 and Ets-1 play an essential role in transcription of FKBP12.6 and FKBP12, respectively.
AB - Cyclic ADP-ribose (cADPR) induces the release of Ca2+ from microsomes of pancreatic islets for insulin secretion. It has been demonstrated that cADPR binds to FK506-binding protein 12.6 (FKBP 12.6) on rat islet ryanodine receptor and that the binding of cADPR to FKBP12.6 frees the ryanodine receptor from FKBP12.6, causing it to release Ca2+ [Noguchi, N., Takasawa, S., Nata, K., Tohgo, A., Kato, I., Ikehata, F., Yonekura, H., Okamoto, H., 1997. Cyclic ADP-ribose binds to FK506-binding protein to release Ca 2+ from islet microsomes. J. Biol. Chem. 272, 3133-3136.]. In this study, we cloned, characterized the structural organization of the human FKBP12.6, which is highly homologous to human FKBP12, and analyzed the promoters for FKBP12.6 and FKBP12. Human FKBP12.6 gene spanned about 16 kb in length and consisted of four exons and three introns. The positions of exon-intron junction of the FKBP12.6 gene were perfectly matched with those of FKBP12 gene except that FKBP12 has an additional exon, exon V, to code exclusively for 3′-UTR. Fluorescence in situ hybridization revealed that the FKBP12.6 gene was located on chromosome 2 p21-23, which is different from the locus (chromosome 20 p13) of the FKBP12 gene. Reporter gene analyses revealed that the regions of - 58 ∼ - 24 of FKBP12.6 and - 106 ∼ - 79 of FKBP12 are important for promoter activities. The promoters contain a consensus transcription factor binding sequence for Sp family in FKBP12.6 and Ets-1 in FKBP12. Electrophoretic mobility shift assays showed that nuclear proteins bind to the promoters. The DNA/protein complex on FKBP12.6 promoter was competed out by Sp1 consensus probe and the complex was supershifted by anti-Sp3 antibodies. On the other hand, the DNA/protein complex on FKBP12 promoter was competed out by Ets-1 consensus probe but not by its mutant probe, indicating that Sp3 and Ets-1 play an essential role in transcription of FKBP12.6 and FKBP12, respectively.
KW - Cyclic ADP-ribose
KW - FKBP12
KW - FKBP12.6
KW - Gene structure
KW - Promoter assay
KW - Ryanodine receptor
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U2 - 10.1016/j.gene.2005.07.004
DO - 10.1016/j.gene.2005.07.004
M3 - Article
C2 - 16122887
AN - SCOPUS:26444434268
VL - 360
SP - 55
EP - 64
JO - Gene
JF - Gene
SN - 0378-1119
IS - 1
ER -