Parathyroid hormone (PTH) suppresses rat PTH/PTH-related protein receptor gene promoter.
Parathyroid hormone (PTH) regulates osteoblasts via a G protein-linked PTH/PTH-related protein (PTHrP) receptor. PTH effects on PTH/PTHrP receptor gene expression were studied in UMR 106 osteoblast-like cells. In heterogeneous nuclear RNA and Northern analysis, PTH suppressed PTH/PTHrP receptor transcription. We cloned the 7-kb promoter region of the rat PTH/PTHrP receptor gene and transiently transfected chimeric deletion constructs containing the 5'-flanking region and the luciferase gene into UMR 106 cells. In transfected cells the minimal region for basal promoter activity was between positions -128 and +103. The 5'-flanking region of exon U1 contained several putative-binding sites for Sp1 and the myc-associated zinc finger protein (MAZ). The minimal PTH-suppressive region (PTHSR) was between +1 and +25 in exon U1, but the 5'-flanking region or Sp1 and MAZ-binding sites also were required for PTH-mediated repression. By gel mobility shift assay PTH markedly decreased binding of PTHSR-protein complex in UMR 106 cells. The mutation experiments showed that the most critical sequence for the repression of PTH was 5'-GGGGGAGGGGAG-3' (+1 to +12) of PTHSR. This represents the first characterization of a PTH-suppressive region of the PTH/PTHrP receptor gene in rat.
Kawane
T
T
Department of Biochemistry, Ohu University School of Dentistry, Koriyama 963-8611, Japan.
Mimura
J
J
Fujii-Kuriyama
Y
Y
Horiuchi
N
N
D000818:Animals; D001483:Base Sequence; D004247:DNA / Q000032:analysis; D020862:GC Rich Sequence / Q000235:genetics; D020868:Gene Silencing / Q000502:physiology*; D008969:Molecular Sequence Data; D010006:Osteoblasts / Q000502:physiology; D010281:Parathyroid Hormone / Q000502:physiology*; D011401:Promoter Regions, Genetic / Q000235:genetics*; D051381:Rats; D044168:Receptor, Parathyroid Hormone, Type 1; D018016:Receptors, Parathyroid Hormone / Q000235:genetics*; D014407:Tumor Cells, Cultured
D010281:Parathyroid Hormone; D044168:Receptor, Parathyroid Hormone, Type 1; D018016:Receptors, Parathyroid Hormone; D004247:DNA