TY - JOUR
T1 - Cloning of a coproporphyrinogen oxidase promoter regulatory element binding protein
AU - Takahashi, Shinichiro
AU - Furuyama, Kazumichi
AU - Kobayashi, Akira
AU - Taketani, Shigeru
AU - Harigae, Hideo
AU - Yamamoto, Masayuki
AU - Igarashi, Kazuhiko
AU - Sasaki, Takeshi
AU - Hayashi, Norio
N1 - Funding Information:
We thank Dr. S. Sassa for helpful comments, Dr. E. Ito for providing λgt11 K562 cDNA library, and Dr. J. Mimura for providing pX4TK-Luc reporter plasmid. We also thank Drs. K. Meguro, J. Kameoka, and T. Nagai for stimulating discussions. This work is supported in part by Grants-in-Aid from the Ministry of Education, Science, Sports, and Culture, Japan (to A.K., K.I., H.H., M.Y., T.S., and N.H.)
PY - 2000/7/5
Y1 - 2000/7/5
N2 - Coproporphyrinogen oxidase [CPO] gene promoter regulatory element (CPRE) plays an important role in CPO gene regulation. To isolate a CPRE binding protein, we performed Southwestern screening of K562 cDNA expression library using CPRE as a probe and isolated a cDNA clone which encoded a novel protein, Klp1 (K562 cell-derived leucine-zipper-like protein 1). Klp1 mRNA was highly expressed in K562 cells, HeLa cells, and brain as a single transcript (1.4 kb). Gel mobility shift assays revealed that Klp1 specifically binds to CPRE. Computational analysis revealed that Klp1 has a leucine-zipper-like structure, a Leu-X-X-Leu-Leu motif, and a putative nuclear localization signal in the basic amino acid rich region. Transfection of the Klp1 expression vector into THP-1 cells resulted in transcriptional activation of a reporter construct containing CPRE. These results indicate that Klp1 is a DNA sequence-specific transcription factor that regulates gene expression of genes that contain CPRE in their regulatory region. (C) 2000 Academic Press.
AB - Coproporphyrinogen oxidase [CPO] gene promoter regulatory element (CPRE) plays an important role in CPO gene regulation. To isolate a CPRE binding protein, we performed Southwestern screening of K562 cDNA expression library using CPRE as a probe and isolated a cDNA clone which encoded a novel protein, Klp1 (K562 cell-derived leucine-zipper-like protein 1). Klp1 mRNA was highly expressed in K562 cells, HeLa cells, and brain as a single transcript (1.4 kb). Gel mobility shift assays revealed that Klp1 specifically binds to CPRE. Computational analysis revealed that Klp1 has a leucine-zipper-like structure, a Leu-X-X-Leu-Leu motif, and a putative nuclear localization signal in the basic amino acid rich region. Transfection of the Klp1 expression vector into THP-1 cells resulted in transcriptional activation of a reporter construct containing CPRE. These results indicate that Klp1 is a DNA sequence-specific transcription factor that regulates gene expression of genes that contain CPRE in their regulatory region. (C) 2000 Academic Press.
KW - CPO
KW - CPRE
KW - K562 cells
KW - Klp1
KW - Leu-X-X-Leu-Leu motif
KW - Leucine-zipper-like protein
KW - Southwestern screening
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U2 - 10.1006/bbrc.2000.2991
DO - 10.1006/bbrc.2000.2991
M3 - Article
C2 - 10873651
AN - SCOPUS:0033873204
VL - 273
SP - 596
EP - 602
JO - Biochemical and Biophysical Research Communications
JF - Biochemical and Biophysical Research Communications
SN - 0006-291X
IS - 2
ER -