Characterisation of the Mouse Vasoactive Intestinal Peptide Receptor Type 2 Gene, Vipr2, and Identification of a Polymorphic LINE-1-like Sequence That Confers Altered Promoter Activity

The VPAC2 receptor is a seven transmembrane spanning G protein‐coupled receptor for two neuropeptides, vasoactive intestinal peptide (VIP) and pituitary adenylate cyclase‐activating polypeptide (PACAP). It has a distinct tissue‐specific, developmental and inducible expression that underlies an impor...

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Veröffentlicht in:Journal of neuroendocrinology 2007-01, Vol.19 (1), p.14-25
Hauptverfasser: Steel, G., Lutz, E. M.
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description The VPAC2 receptor is a seven transmembrane spanning G protein‐coupled receptor for two neuropeptides, vasoactive intestinal peptide (VIP) and pituitary adenylate cyclase‐activating polypeptide (PACAP). It has a distinct tissue‐specific, developmental and inducible expression that underlies an important neuroendocrine role. Here, we report the characterisation of the gene that encodes the mouse VPAC2 receptor (Vipr2), localisation of the transcriptional start site and functional analysis of the promoter region. The Vipr2 gene contains 12 introns within its protein‐coding region and spans 68.6 kb. Comparison of the 5′ untranslated region sequences for cloned 5′‐RACE products amplified from different tissues showed they all were contained within the same exon, with the longest extending 111 bp upstream of the ATG start site. Functional analysis of the 3.2‐kb 5′‐flanking region using sequentially deleted sequences cloned into a luciferase gene reporter vector revealed that this region is active as a promoter in mouse AtT20 D16:16 and rat GH4C1 cell lines. The core promoter is located within a 180‐bp GC‐rich region proximal to the ATG start codon and contains potential binding sites for Sp1 and AP2, but no TATA‐box. Further upstream, in two out of three mice strains examined, we have discovered a 496‐bp polymorphic DNA sequence that bears a significant identity to mouse LINE‐1 DNA. Comparison of the promoter activity between luciferase reporter gene constructs derived from the BALB/c (which contains this sequence) and C57BL/6J (which lacks this sequence) Vipr2 promoter regions has shown three‐fold difference in luciferase gene activity when expressed in mouse AtT20 D16:16 and αT3‐1 cells, but not when expressed in the rat GH4C1 cells or in COS 7 cells. Our results suggest that the mouse Vipr2 gene may be differentially active in different mouse strains, depending on the presence of this LINE‐1‐like sequence in the promoter region.
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M.</creator><creatorcontrib>Steel, G. ; Lutz, E. M.</creatorcontrib><description>The VPAC2 receptor is a seven transmembrane spanning G protein‐coupled receptor for two neuropeptides, vasoactive intestinal peptide (VIP) and pituitary adenylate cyclase‐activating polypeptide (PACAP). It has a distinct tissue‐specific, developmental and inducible expression that underlies an important neuroendocrine role. Here, we report the characterisation of the gene that encodes the mouse VPAC2 receptor (Vipr2), localisation of the transcriptional start site and functional analysis of the promoter region. The Vipr2 gene contains 12 introns within its protein‐coding region and spans 68.6 kb. Comparison of the 5′ untranslated region sequences for cloned 5′‐RACE products amplified from different tissues showed they all were contained within the same exon, with the longest extending 111 bp upstream of the ATG start site. Functional analysis of the 3.2‐kb 5′‐flanking region using sequentially deleted sequences cloned into a luciferase gene reporter vector revealed that this region is active as a promoter in mouse AtT20 D16:16 and rat GH4C1 cell lines. The core promoter is located within a 180‐bp GC‐rich region proximal to the ATG start codon and contains potential binding sites for Sp1 and AP2, but no TATA‐box. Further upstream, in two out of three mice strains examined, we have discovered a 496‐bp polymorphic DNA sequence that bears a significant identity to mouse LINE‐1 DNA. Comparison of the promoter activity between luciferase reporter gene constructs derived from the BALB/c (which contains this sequence) and C57BL/6J (which lacks this sequence) Vipr2 promoter regions has shown three‐fold difference in luciferase gene activity when expressed in mouse AtT20 D16:16 and αT3‐1 cells, but not when expressed in the rat GH4C1 cells or in COS 7 cells. 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M.</creatorcontrib><title>Characterisation of the Mouse Vasoactive Intestinal Peptide Receptor Type 2 Gene, Vipr2, and Identification of a Polymorphic LINE-1-like Sequence That Confers Altered Promoter Activity</title><title>Journal of neuroendocrinology</title><addtitle>J Neuroendocrinol</addtitle><description>The VPAC2 receptor is a seven transmembrane spanning G protein‐coupled receptor for two neuropeptides, vasoactive intestinal peptide (VIP) and pituitary adenylate cyclase‐activating polypeptide (PACAP). It has a distinct tissue‐specific, developmental and inducible expression that underlies an important neuroendocrine role. Here, we report the characterisation of the gene that encodes the mouse VPAC2 receptor (Vipr2), localisation of the transcriptional start site and functional analysis of the promoter region. The Vipr2 gene contains 12 introns within its protein‐coding region and spans 68.6 kb. Comparison of the 5′ untranslated region sequences for cloned 5′‐RACE products amplified from different tissues showed they all were contained within the same exon, with the longest extending 111 bp upstream of the ATG start site. Functional analysis of the 3.2‐kb 5′‐flanking region using sequentially deleted sequences cloned into a luciferase gene reporter vector revealed that this region is active as a promoter in mouse AtT20 D16:16 and rat GH4C1 cell lines. The core promoter is located within a 180‐bp GC‐rich region proximal to the ATG start codon and contains potential binding sites for Sp1 and AP2, but no TATA‐box. Further upstream, in two out of three mice strains examined, we have discovered a 496‐bp polymorphic DNA sequence that bears a significant identity to mouse LINE‐1 DNA. 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M.</au><format>journal</format><genre>article</genre><ristype>JOUR</ristype><atitle>Characterisation of the Mouse Vasoactive Intestinal Peptide Receptor Type 2 Gene, Vipr2, and Identification of a Polymorphic LINE-1-like Sequence That Confers Altered Promoter Activity</atitle><jtitle>Journal of neuroendocrinology</jtitle><addtitle>J Neuroendocrinol</addtitle><date>2007-01</date><risdate>2007</risdate><volume>19</volume><issue>1</issue><spage>14</spage><epage>25</epage><pages>14-25</pages><issn>0953-8194</issn><eissn>1365-2826</eissn><abstract>The VPAC2 receptor is a seven transmembrane spanning G protein‐coupled receptor for two neuropeptides, vasoactive intestinal peptide (VIP) and pituitary adenylate cyclase‐activating polypeptide (PACAP). It has a distinct tissue‐specific, developmental and inducible expression that underlies an important neuroendocrine role. Here, we report the characterisation of the gene that encodes the mouse VPAC2 receptor (Vipr2), localisation of the transcriptional start site and functional analysis of the promoter region. The Vipr2 gene contains 12 introns within its protein‐coding region and spans 68.6 kb. Comparison of the 5′ untranslated region sequences for cloned 5′‐RACE products amplified from different tissues showed they all were contained within the same exon, with the longest extending 111 bp upstream of the ATG start site. Functional analysis of the 3.2‐kb 5′‐flanking region using sequentially deleted sequences cloned into a luciferase gene reporter vector revealed that this region is active as a promoter in mouse AtT20 D16:16 and rat GH4C1 cell lines. The core promoter is located within a 180‐bp GC‐rich region proximal to the ATG start codon and contains potential binding sites for Sp1 and AP2, but no TATA‐box. Further upstream, in two out of three mice strains examined, we have discovered a 496‐bp polymorphic DNA sequence that bears a significant identity to mouse LINE‐1 DNA. Comparison of the promoter activity between luciferase reporter gene constructs derived from the BALB/c (which contains this sequence) and C57BL/6J (which lacks this sequence) Vipr2 promoter regions has shown three‐fold difference in luciferase gene activity when expressed in mouse AtT20 D16:16 and αT3‐1 cells, but not when expressed in the rat GH4C1 cells or in COS 7 cells. Our results suggest that the mouse Vipr2 gene may be differentially active in different mouse strains, depending on the presence of this LINE‐1‐like sequence in the promoter region.</abstract><cop>Oxford, UK</cop><pub>Blackwell Publishing Ltd</pub><pmid>17184482</pmid><doi>10.1111/j.1365-2826.2006.01498.x</doi><tpages>12</tpages><oa>free_for_read</oa></addata></record>
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identifier ISSN: 0953-8194
ispartof Journal of neuroendocrinology, 2007-01, Vol.19 (1), p.14-25
issn 0953-8194
1365-2826
language eng
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source MEDLINE; Wiley Journals
subjects Animals
Base Sequence
Biological and medical sciences
Cells, Cultured
Chlorocebus aethiops
COS Cells
Fundamental and applied biological sciences. Psychology
G protein-coupled receptor
gene
LINE-1
Long Interspersed Nucleotide Elements
Mice
Mice, Inbred BALB C
Mice, Inbred C57BL
Molecular Sequence Data
Original
pituitary adenylate cyclase activating polypeptide (PACAP)
Polymorphism, Genetic
Promoter Regions, Genetic
Rats
Receptors, Vasoactive Intestinal Peptide, Type II - genetics
Sequence Homology, Nucleic Acid
Somatotrophs
vasoactive intestinal peptide (VIP)
Vertebrates: endocrinology
title Characterisation of the Mouse Vasoactive Intestinal Peptide Receptor Type 2 Gene, Vipr2, and Identification of a Polymorphic LINE-1-like Sequence That Confers Altered Promoter Activity
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