Human Vasoactive Intestinal Peptide1 Receptors Expressed by Stable Transfectants Couple to 2 Distinct Signaling Pathways

Sreedharan, Sunil P.; Patel, Dhruv; Xia, Menghang; Ichikawa, Shintaro; Goetzl, Edward J.

doi:10.1006/bbrc.1994.2160

Cited by 51 publications

(29 citation statements)

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“…2) followed the consensus GT-AG rule (11). Both type I VIP and type I PACAP receptors can positively couple to concurrent increases in intracellular cAMP and free Ca2+, presumably through association with different G proteins (12,13). Five spliced variants of the type I PACAP receptor cDNA have been identified with insertions at the C terminus of the third cytoplasmic loop connecting transmembrane segments V and VI, and these variants appeared to affect receptor coupling to adenylyl cyclase and phospholipase C stimulation (13).…”

Section: Resultsmentioning

confidence: 99%

Structure, expression, and chromosomal localization of the type I human vasoactive intestinal peptide receptor gene.

Sreedharan

Huang

Cheung

et al. 1995

Proc. Natl. Acad. Sci. U.S.A.

148

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Vasoactive intestinal peptide (VIP) and other members of the pituitary adenylyl cyclase-activating peptide (PACAP) and secretin neuroendocrine peptide family are recognized with specificity by related G protein-coupled receptors. We report here the cloning, characterization, and chromosomal location of the gene encoding the human type I VIP receptor (HVR1), also termed the type II PACAP receptor. The gene spans -22 kb and is composed of 13 exons ranging from 42 to 1400 bp and 12 introns ranging from 0.3 to 6.1 kb. Primer extension analysis with poly(A)+ RNA from human HT29 colonic adenocarcinoma cells indicated that the transcription initiation site is located at position -110 upstream of the first nucleotide (+1) of the translation start codon, and 75 nt downstream of a consensus CCAAT-box motif. The G+C-rich 5' flanking region contains potential binding sites for several nuclear factors, including Spl, AP2, ATF, interferon regulatory factor 1, NF-IL6, acute-phase response factor, and NF-KcB. The HVRI gene is expressed selectively in human tissues with a relative prevalence of lung > prostate > peripheral blood leukocytes, liver, brain, small intestine > colon, heart, spleen > placenta, kidney, thymus, testis. Fluorescence in situ hybridization localized the HVR1 gene to the short arm of human chromosome 3 (3p22), in a region associated with small-cell lung cancer.The 28-aa vasoactive intestinal peptide (VIP) is structurally related to other members of a family of peptide neuroendocrine mediators that include pituitary adenylyl cyclaseactivating peptide (PACAP), secretin, glucagon, calcitonin, and parathyroid hormone. VIP has potent relaxing effects on nonvascular and vascular smooth muscle, enhancing blood flow. It also regulates water and ion flux from lung and intestinal epithelia, promotes neuronal growth and survival, and modulates many immune functions (1-3).Specific high-affinity receptors for VIP are found on distinct subsets of neural, respiratory, gastrointestinal, and immune cells (1-3). We previously cloned a cDNA encoding a human high-affinity type I VIP receptor (HVR1), also termed type II PACAP receptor, from HT29 colonic adenocarcinoma cells and human lung tissue (4). The 3-kb cDNA insert encoded a seven-transmembrane-domain HVR1 protein of 457 aa, with a deduced molecular mass of 52 kDa, that was most homologous to other members of the PACAP and secretin G proteincoupled receptor family and had a similar binding affinity for both VIP and PACAP (4 Analysis of Introns. The size of introns in the HVR1 gene was determined by polymerase chain reaction (9) of genomic clones with specific primers derived from exon sequences, for 35 cycles of 30 sec at 94°C, 2 min at 55°C, and 6 min at 72°C, using Taq polymerase (GIBCO/BRL). The reaction products were analyzed by electrophoresis in 1% agarose gels.Primer Extension. Two micrograms of poly(A)+ RNA from HT29 cells was hybridized overnight with 50,000 cpm of a 32P-end-labeled primer (5'-ACTTGGCGGGCGCATG-GTCT-3') which spans the cDNA translation sta...

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Section: Resultsmentioning

confidence: 99%

Structure, expression, and chromosomal localization of the type I human vasoactive intestinal peptide receptor gene.

Sreedharan

Huang

Cheung

et al. 1995

Proc. Natl. Acad. Sci. U.S.A.

148

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“…Ruiz-Velasco and Ikeda (38) found they could activate Kir3.1ϩ3.2 and Kir3.1ϩ3.4 channels heterologously expressed in superior cervical ganglion neurons through the vasoactive intestinal peptide (VIP) receptor. VIP does not exclusively stimulate G s -coupled receptors: it has been shown to couple to G␣ i1/2 (40) and also to elevate intracellular Ca 2ϩ levels (41). The VIP-mediated effects were enhanced by pre-treatment with PTx, leading to the authors' suggestion that the uncoupling of PTx-sensitive G proteins allowed more PTx-insensitive G proteins to activate the channels.…”

Section: Discussionmentioning

confidence: 99%

The G Protein α Subunit Has a Key Role in Determining the Specificity of Coupling to, but Not the Activation of, G Protein-gated Inwardly Rectifying K+ Channels

Leaney¹,

Milligan²,

Tinker³

2000

Journal of Biological Chemistry

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In neuronal and atrial tissue, G protein-gated inwardly rectifying K ؉ channels (Kir3.x family) are responsible for mediating inhibitory postsynaptic potentials and slowing the heart rate. They are activated by G␤␥ dimers released in response to the stimulation of receptors coupled to inhibitory G proteins of the G i/o family but not receptors coupled to the stimulatory G protein G s . We have used biochemical, electrophysiological, and molecular biology techniques to examine this specificity of channel activation. In this study we have succeeded in reconstituting such specificity in an heterologous expression system stably expressing a cloned counterpart of the neuronal channel (Kir3.1 and Kir3.2A heteromultimers). The use of pertussis toxinresistant G protein ␣ subunits and chimeras between G i1 and G s indicate a central role for the G protein ␣ subunits in determining receptor specificity of coupling to, but not activation of, G protein-gated inwardly rectifying K ؉ channels.G protein-gated, inwardly rectifying K ϩ channels were first identified in atrial myocytes where they were shown to be activated by acetylcholine at muscarinic m 2 receptors (1), and stimulation of this current is responsible for slowing of the heart rate in response to vagal nerve stimulation. It was subsequently shown that this activation was sensitive to pertussis toxin (PTx), 1 implicating the family of G i/o proteins (2-4). It is now apparent that G protein-gated inwardly rectifying K ϩ currents are present in many neuronal cell types, and they are involved in the postsynaptic inhibitory effects of stimulating G i/o -coupled receptors such as GABA B and adenosine A 1 (5).Cloning efforts from a number of laboratories have revealed the molecular counterparts of these currents (6 -10). The channel is a heteromultimer of members of the inwardly rectifying Kir3.x family of potassium channels. Co-expression of Kir3.1 with Kir3.2, Kir3.3, or Kir3.4 results in currents that show many of the basic characteristics of the native channels in neurons and atria (11-13). Homomultimers of splice variants of Kir3.2 may also form in certain neuronal regions (14).Channel activation of these currents in native tissues and of the cloned counterparts in heterologous expression systems was shown to be membrane-delimited (4, 15), involving a direct interaction with the G protein ␤␥ dimer (16). This finding was initially controversial, but there are now numerous lines of evidence that point to the involvement of the ␤␥ subunit as the direct channel activator (17). Recent studies have focused on domains and residues involved in G protein ␤␥ dimer binding (18 -24) and the cell biology of channel biogenesis (25-27).The question remains as to how receptor specificity is achieved and what are the key determinants involved in this. In native tissues only G i/o -coupled receptors, and not G s -coupled receptors, activate these channels, and yet both should liberate free G protein ␤␥ upon receptor stimulation. Receptor selectivity does not lie at the level of the G...

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“…Although only a single population of binding sites had been identified in wt HT-29 cells , VIP can be covalently cross-linked to two distinct proteins in normal human colonic epithelial membranes, in both a specific and GTP-sensitive manner . Alternatively, the same receptor might be coupled to different downstream pathways dependent on its subcellular localization, for example to different second messenger systems (Sreedharan et al, 1994).…”

Section: Pro34mentioning

confidence: 99%

The functional investigation of a human adenocarcinoma cell line, stably transfected with the neuropeptide Y Y₁ receptor

Holliday

Cox

1996

British J Pharmacology

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1 The human adenocarcinoma cell line, HT-29, has been stably transfected with the cDNA sequence for the rat neuropeptide Y (NPY) Y1 receptor, and three Y1 clones (Y1-4, Y1-7 and Y1 -16) have been isolated which express high levels of specific ['251 6 HT-29 clones stably expressing the Y1 receptor therefore show responses to PYY and its analogues that are characteristic of that subtype, and the Y1-7 clone in particular will be useful in the assessment of novel Y1-specific drugs. This approach will also allow the functional study of NPY Y1 receptors with selected mutations.

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Human Vasoactive Intestinal Peptide1 Receptors Expressed by Stable Transfectants Couple to 2 Distinct Signaling Pathways

Cited by 51 publications

References 0 publications

Structure, expression, and chromosomal localization of the type I human vasoactive intestinal peptide receptor gene.

Structure, expression, and chromosomal localization of the type I human vasoactive intestinal peptide receptor gene.

The G Protein α Subunit Has a Key Role in Determining the Specificity of Coupling to, but Not the Activation of, G Protein-gated Inwardly Rectifying K+ Channels

The functional investigation of a human adenocarcinoma cell line, stably transfected with the neuropeptide Y Y₁ receptor

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Human Vasoactive Intestinal Peptide1 Receptors Expressed by Stable Transfectants Couple to 2 Distinct Signaling Pathways

Cited by 51 publications

References 0 publications

Structure, expression, and chromosomal localization of the type I human vasoactive intestinal peptide receptor gene.

Structure, expression, and chromosomal localization of the type I human vasoactive intestinal peptide receptor gene.

The G Protein α Subunit Has a Key Role in Determining the Specificity of Coupling to, but Not the Activation of, G Protein-gated Inwardly Rectifying K+ Channels

The functional investigation of a human adenocarcinoma cell line, stably transfected with the neuropeptide Y Y1 receptor

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The functional investigation of a human adenocarcinoma cell line, stably transfected with the neuropeptide Y Y₁ receptor