Tissue-specific transcription start sites and alternative splicing of the parathyroid hormone (PTH)/PTH-related peptide (PTHrP) receptor gene: A new PTH/PTHrP receptor splice variant that lacks the signal peptide

被引:60
作者
Joun, H
Lanske, B
Karperien, M
Qian, F
Defize, L
AbouSamra, A
机构
[1] MASSACHUSETTS GEN HOSP,ENDOCRINE UNIT,BOSTON,MA 02114
[2] NETHERLANDS INST DEV BIOL,UTRECHT,NETHERLANDS
关键词
D O I
10.1210/en.138.4.1742
中图分类号
R5 [内科学];
学科分类号
1002 ; 100201 ;
摘要
The PTH/PTHrP receptor gene is expressed in bone and kidney as well as in many other tissues. Using primer extension followed by rapid cloning of amplified complementary DNA ends, we have isolated new PTH/PTHrP receptor complementary DNAs with different splicing patterns and have characterized a new upstream transcription start site. Three 5' nontranslated exons, U3, U2 and U1, located 4.8, 2.5, and 1.2 kb upstream of the exon that encodes the putative signal peptide of the classical receptor (exon S), have been characterized. Four types of splicing patterns were recognized. Type I splicing pattern is transcribed from exon U1 and is spliced to exons S and E1; this pattern was found in most tissues tested. Types II, III, and IV splicing patterns are transcribed from exon U3 and have a restricted tissue distribution. Type II splice pattern, containing exons U3, U2, and S and type III splicing pattern, containing exon U3, U2, and El (skipping exon S), was found only in kidney. Type IV splice pattern, containing exon U3 and S was found both in kidney and ovary. Because the type III splice variant skips exon S, translation of this splice Variant initiates at a different AUG codon. The type III splice variant was weakly expressed on the cell surface of COS-7 cells, as assessed by double antibody binding assay, and no detectable ligand binding was observed on intact cells. The type III splice variant, however, increased cAMP accumulation in COS-7 cells when challenged with PTH(1-34), PTH(1-84) and hPTHrP(1-36) with EC50s that are similar to those observed in COS-7 cells expressing the type I variant but with a maximum stimulation that was lower than that observed in COS-7 cells expressing the type I variant. These data indicate low levels of cell surface expression of the type III splice variant. Treatment of COS-7 cells with tunicamycin decreased the size of the type I splice variant from a broad band of 85 kDa to a compact band of about 60 kDa. The type III splice variant did not change in size in COS-7 cells treated with tunicamycin, indicating that the type III splice variant did not undergo any glycosylation step. In conclusion, the PTH/PTHrP receptor gene uses alternate promoters in a tissue-specific manner that results in several tissue-specific alternatively spliced transcripts. One of these transcripts, the type III splice variant, is expressed in kidney and lacks the signal peptide.
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页码:1742 / 1749
页数:8
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共 24 条
  • [1] ABOUSAMRA A, 1993, END SOC M LAS VEG NE
  • [2] EXPRESSION CLONING OF A COMMON RECEPTOR FOR PARATHYROID-HORMONE AND PARATHYROID HORMONE-RELATED PEPTIDE FROM RAT OSTEOBLAST-LIKE CELLS - A SINGLE RECEPTOR STIMULATES INTRACELLULAR ACCUMULATION OF BOTH CAMP AND INOSITOL TRISPHOSPHATES AND INCREASES INTRACELLULAR FREE CALCIUM
    ABOUSAMRA, AB
    JUPPNER, H
    FORCE, T
    FREEMAN, MW
    KONG, XF
    SCHIPANI, E
    URENA, P
    RICHARDS, J
    BONVENTRE, JV
    POTTS, JT
    KRONENBERG, HM
    SEGRE, GV
    [J]. PROCEEDINGS OF THE NATIONAL ACADEMY OF SCIENCES OF THE UNITED STATES OF AMERICA, 1992, 89 (07) : 2732 - 2736
  • [3] DOWN-REGULATION OF PARATHYROID (PTH) PTH-RELATED PEPTIDE RECEPTOR IMMUNOREACTIVITY AND PTH BINDING IN OPOSSUM KIDNEY-CELLS BY PTH AND DEXAMETHASONE
    ABOUSAMRA, AB
    GOLDSMITH, PK
    XIE, LY
    JUPPNER, H
    SPIEGEL, AM
    SEGRE, GV
    [J]. ENDOCRINOLOGY, 1994, 135 (06) : 2588 - 2594
  • [4] PROBING THE BIMOLECULAR INTERACTIONS OF PARATHYROID-HORMONE AND THE HUMAN PARATHYROID-HORMONE PARATHYROID HORMONE-RELATED PROTEIN-RECEPTOR .2. CLONING, CHARACTERIZATION, AND PHOTOAFFINITY-LABELING OF THE RECOMBINANT HUMAN RECEPTOR
    ADAMS, AE
    PINES, M
    NAKAMOTO, C
    BEHAR, V
    YANG, QM
    BESSALLE, R
    CHOREV, M
    ROSENBLATT, M
    LEVINE, MA
    SUVA, LJ
    [J]. BIOCHEMISTRY, 1995, 34 (33) : 10553 - 10559
  • [5] FROHMAN MA, 1990, AMPLIFICATIONS, V5, P11
  • [6] HENDERSON JE, 1995, MOL CELL BIOL, V15, P4064
  • [7] JOUN H, 1994, AM SOC BONE MINER S1, V9, pS285
  • [8] A G-PROTEIN LINKED RECEPTOR FOR PARATHYROID-HORMONE AND PARATHYROID-HORMONE RELATED PEPTIDE
    JUPPNER, H
    ABOUSAMRA, AB
    FREEMAN, M
    KONG, XF
    SCHIPANI, E
    RICHARDS, J
    KOLAKOWSKI, LF
    HOCK, J
    POTTS, JT
    KRONENBERG, HM
    SEGRE, GV
    [J]. SCIENCE, 1991, 254 (5034) : 1024 - 1026
  • [9] JUPPNER H, 1988, J BIOL CHEM, V263, P8557
  • [10] EXPRESSION PATTERN OF PARATHYROID-HORMONE PARATHYROID-HORMONE RELATED PEPTIDE RECEPTOR MESSENGER-RNA IN MOUSE POSTIMPLANTATION EMBRYOS INDICATES INVOLVEMENT IN MULTIPLE DEVELOPMENTAL PROCESSES
    KARPERIEN, M
    VANDIJK, TB
    HOEIJMAKERS, T
    CREMERS, F
    ABOUSAMRA, AB
    BOONSTRA, J
    DELAAT, SW
    DEFIZE, LHK
    [J]. MECHANISMS OF DEVELOPMENT, 1994, 47 (01) : 29 - 42