Cloning and characterization of the proximal murine Phex promoter

Shiguang Liu, Rong Guo, Leigh Quarles

Research output: Contribution to journalArticle

23 Citations (Scopus)

Abstract

Phex is an endopetidase that regulates systemic phosphate homeostasis. We investigated Phex gene transcription by cloning and performing functional analysis of the 2736 bp of the 5′ flanking region of the mouse Phex gene containing its promoter. We identified a transcription start site, a consensus Tata-box, and multiple potential cis-acting regulator elements. To determine whether the promoter directs cell-type restricted Phex expression, we transfected full-length and 5′-deleted Phex luciferase reporter constructs into various cell lines. Phex-expressing C5.18 chondrocytes displayed the highest activity of the transfected Phex promoter constructs compared with non-Phex-expressing COS-7 cells, whereas promoter activity was intermediate in ROS 17/2.8 osteoblasts and maturation stage-dependent in MC3T3-E1 osteoblasts. Analysis of sequential 5′-deletion mutants of the Phex promoter in ROS 17/2.8 cells revealed bimodal activity, suggesting that both positive and negative cis-acting regions may be present. The chondrogenic factor SOX9 markedly stimulated Phex promoter activity, whereas Cbfa1, PTH, and 1,25(OH)2D3 had no effect. Our findings are consistent with the predominant expression of Phex in bone and cartilage. Additional studies will be needed to confirm the regulatory regions in the Phex promoter that function in a cell-restricted manner.

Original languageEnglish (US)
Pages (from-to)3987-3995
Number of pages9
JournalEndocrinology
Volume142
Issue number9
DOIs
StatePublished - Jan 1 2001
Externally publishedYes

Fingerprint

Organism Cloning
Osteoblasts
5' Flanking Region
Nucleic Acid Regulatory Sequences
Transcription Initiation Site
COS Cells
Chondrocytes
Luciferases
Genes
Cartilage
Homeostasis
Phosphates
Bone and Bones
Cell Line

All Science Journal Classification (ASJC) codes

  • Endocrinology

Cite this

Cloning and characterization of the proximal murine Phex promoter. / Liu, Shiguang; Guo, Rong; Quarles, Leigh.

In: Endocrinology, Vol. 142, No. 9, 01.01.2001, p. 3987-3995.

Research output: Contribution to journalArticle

Liu, Shiguang ; Guo, Rong ; Quarles, Leigh. / Cloning and characterization of the proximal murine Phex promoter. In: Endocrinology. 2001 ; Vol. 142, No. 9. pp. 3987-3995.
@article{a27dfe2d40df49a794d5dd926dee6057,
title = "Cloning and characterization of the proximal murine Phex promoter",
abstract = "Phex is an endopetidase that regulates systemic phosphate homeostasis. We investigated Phex gene transcription by cloning and performing functional analysis of the 2736 bp of the 5′ flanking region of the mouse Phex gene containing its promoter. We identified a transcription start site, a consensus Tata-box, and multiple potential cis-acting regulator elements. To determine whether the promoter directs cell-type restricted Phex expression, we transfected full-length and 5′-deleted Phex luciferase reporter constructs into various cell lines. Phex-expressing C5.18 chondrocytes displayed the highest activity of the transfected Phex promoter constructs compared with non-Phex-expressing COS-7 cells, whereas promoter activity was intermediate in ROS 17/2.8 osteoblasts and maturation stage-dependent in MC3T3-E1 osteoblasts. Analysis of sequential 5′-deletion mutants of the Phex promoter in ROS 17/2.8 cells revealed bimodal activity, suggesting that both positive and negative cis-acting regions may be present. The chondrogenic factor SOX9 markedly stimulated Phex promoter activity, whereas Cbfa1, PTH, and 1,25(OH)2D3 had no effect. Our findings are consistent with the predominant expression of Phex in bone and cartilage. Additional studies will be needed to confirm the regulatory regions in the Phex promoter that function in a cell-restricted manner.",
author = "Shiguang Liu and Rong Guo and Leigh Quarles",
year = "2001",
month = "1",
day = "1",
doi = "10.1210/endo.142.9.8403",
language = "English (US)",
volume = "142",
pages = "3987--3995",
journal = "Endocrinology",
issn = "0013-7227",
publisher = "The Endocrine Society",
number = "9",

}

TY - JOUR

T1 - Cloning and characterization of the proximal murine Phex promoter

AU - Liu, Shiguang

AU - Guo, Rong

AU - Quarles, Leigh

PY - 2001/1/1

Y1 - 2001/1/1

N2 - Phex is an endopetidase that regulates systemic phosphate homeostasis. We investigated Phex gene transcription by cloning and performing functional analysis of the 2736 bp of the 5′ flanking region of the mouse Phex gene containing its promoter. We identified a transcription start site, a consensus Tata-box, and multiple potential cis-acting regulator elements. To determine whether the promoter directs cell-type restricted Phex expression, we transfected full-length and 5′-deleted Phex luciferase reporter constructs into various cell lines. Phex-expressing C5.18 chondrocytes displayed the highest activity of the transfected Phex promoter constructs compared with non-Phex-expressing COS-7 cells, whereas promoter activity was intermediate in ROS 17/2.8 osteoblasts and maturation stage-dependent in MC3T3-E1 osteoblasts. Analysis of sequential 5′-deletion mutants of the Phex promoter in ROS 17/2.8 cells revealed bimodal activity, suggesting that both positive and negative cis-acting regions may be present. The chondrogenic factor SOX9 markedly stimulated Phex promoter activity, whereas Cbfa1, PTH, and 1,25(OH)2D3 had no effect. Our findings are consistent with the predominant expression of Phex in bone and cartilage. Additional studies will be needed to confirm the regulatory regions in the Phex promoter that function in a cell-restricted manner.

AB - Phex is an endopetidase that regulates systemic phosphate homeostasis. We investigated Phex gene transcription by cloning and performing functional analysis of the 2736 bp of the 5′ flanking region of the mouse Phex gene containing its promoter. We identified a transcription start site, a consensus Tata-box, and multiple potential cis-acting regulator elements. To determine whether the promoter directs cell-type restricted Phex expression, we transfected full-length and 5′-deleted Phex luciferase reporter constructs into various cell lines. Phex-expressing C5.18 chondrocytes displayed the highest activity of the transfected Phex promoter constructs compared with non-Phex-expressing COS-7 cells, whereas promoter activity was intermediate in ROS 17/2.8 osteoblasts and maturation stage-dependent in MC3T3-E1 osteoblasts. Analysis of sequential 5′-deletion mutants of the Phex promoter in ROS 17/2.8 cells revealed bimodal activity, suggesting that both positive and negative cis-acting regions may be present. The chondrogenic factor SOX9 markedly stimulated Phex promoter activity, whereas Cbfa1, PTH, and 1,25(OH)2D3 had no effect. Our findings are consistent with the predominant expression of Phex in bone and cartilage. Additional studies will be needed to confirm the regulatory regions in the Phex promoter that function in a cell-restricted manner.

UR - http://www.scopus.com/inward/record.url?scp=0034878896&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=0034878896&partnerID=8YFLogxK

U2 - 10.1210/endo.142.9.8403

DO - 10.1210/endo.142.9.8403

M3 - Article

VL - 142

SP - 3987

EP - 3995

JO - Endocrinology

JF - Endocrinology

SN - 0013-7227

IS - 9

ER -