Construction and function of a recombinant adenovirus encoding a human aquaporin 1-green fluorescent protein fusion product

Hoque, A. T. M. Shamsul; Liu, X; Kagami, Hideaki; Swaim, W.D.; Wellner, Robert B.; O’Connell, Brian; Ambudkar, Indu S.; Baum, Bruce J.

doi:10.1038/sj.cgt.7700146

Cited by 15 publications

(16 citation statements)

References 30 publications

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“…Aquaporins tagged with a fluorescent protein at the N terminus are known to retain their functional activity (19,20). The fluorescence of the fusion protein indicated its location in the plasma membrane (Fig.…”

Section: Expression Of Aqp1 and Yfp-aqp1 In Hek293mentioning

confidence: 99%

Aquaporin-1, Nothing but a Water Channel

Tsunoda

Wiesner

Lorenz

et al. 2004

Journal of Biological Chemistry

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Aquaporin-1 (AQP1) is a membrane channel that allows rapid water movement driven by a transmembrane osmotic gradient. It was claimed to have a secondary function as a cyclic nucleotide-gated ion channel. However, upon reconstitution into planar bilayers, the ion channel exhibited a 10-fold lower single channel conductance than in Xenopus oocytes and a 100-fold lower open probability (<10 ؊6 ) of doubtful physiological significance (Saparov, S. M., Kozono, D., Rothe, U., Agre, P., and Pohl, P. (2001) J. Biol. Chem. 276, 31515-31520). Investigating AQP1 expressed in human embryonic kidney cells, we now have shown that the discrepancy is not due to alterations of AQP1 properties upon reconstitution into bilayers but rather to regulatory processes of the oocyte expression system that may have been misinterpreted as AQP1 ion channel activity. As confirmed by laser scanning reflection microscopy, from 0.8 to 1.4 ؋ 10 6 AQP1 copies/cell contributed to osmotic cell swelling. The proper plasma membrane localization was confirmed by observing the fluorescence of the N-terminal yellow fluorescent protein tag. Whole-cell patch clamp experiments of wild type or tagged AQP1-expressing cells revealed that neither cGMP nor cAMP mediated ion channel activity. The lack of significant CNG ion channel activity rules out a secondary role of AQP1 water channels in cellular signal transduction.

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Section: Expression Of Aqp1 and Yfp-aqp1 In Hek293mentioning

confidence: 99%

Aquaporin-1, Nothing but a Water Channel

Tsunoda

Wiesner

Lorenz

et al. 2004

Journal of Biological Chemistry

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“…To determine whether the induction of REG Iα mRNA was caused by the activation of transcription, a 1216-base pair fragment containing 1190-base pair of the promoter region (−1190∼+26) of the human REG Iα gene [73] was fused to the luciferase reporter gene of pGL3-Basic vector and transfected into human NS-SV-DC and rat A5 [74,75] salivary ductal cells using the Lipofectamine™ 2000 reagent. Six hours after transfection, cells were incubated with IL-6 or IL-8 for 24 h, and cell extracts were prepared for luciferase assay.…”

Section: Activation Of Reg Iα Gene Promoter By Il-6mentioning

confidence: 99%

Significance of Interleukin-6/STAT Pathway for the Gene Expression of REG Iα, a New Autoantigen in Sjögren’s Syndrome Patients, in Salivary Duct Epithelial Cells

Fujimura

Fujimoto

Itaya‐Hironaka

et al. 2016

Clinic Rev Allerg Immunol

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The regenerating gene, Reg, was originally isolated from a rat regenerating islet complementary DNA (cDNA) library, and its human homologue was named REG Iα. Recently, we reported that REG Iα messenger RNA (mRNA), as well as its product, was overexpressed in ductal epithelial cells in the salivary glands of Sjögren's syndrome patients. Furthermore, autoantibodies against REG Iα were found in the sera of Sjögren's syndrome patients, and the patients who were positive for the anti-REG Iα antibody showed significantly lower saliva secretion than antibody-negative patients. We found the mechanism of REG Iα induction in salivary ductal epithelial cells. Reporter plasmid containing REG Iα promoter (-1190/+26) upstream of a luciferase gene was introduced into human NS-SV-DC and rat A5 salivary ductal cells. The cells were treated with several cytokines (interleukin (IL)-6, IL-8, etc.), upregulated in Sjögren's syndrome salivary ducts, and the transcriptional activity was measured. IL-6 stimulation significantly enhanced the REG Iα promoter activity in both cells. Deletion analysis revealed that the -141∼-117 region of the REG Iα gene was responsible for the promoter activation by IL-6, which contains a consensus sequence for signal transducer and activator of transcription (STAT) binding. The introduction of small interfering RNA for human STAT3 abolished IL-6-induced REG Iα transcription. These results indicated that IL-6 stimulation induced REG Iα transcription through STAT3 activation and binding to the REG Iα promoter in salivary ductal cells. This dependence of REG Iα induction upon IL-6/STAT in salivary duct epithelial cells may play an important role in the pathogenesis/progression of Sjögren's syndrome.

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“…To determine whether the increase of REG Iα mRNA was caused by the activation of transcription, a 1216-bp fragment containing 1190-bp of the promoter region of the human REG Iα gene was fused to the luciferase gene and transfected into human NS-SV-DC and rat A5 [86,87] salivary ductal cells. We found that IL-6 stimulation significantly enhanced the REG Iα promoter activity not only in NS-SV-DC cells but also in A5 cells.…”

Section: Gene Expression Of Reg Iα As An Auto-antigenmentioning

confidence: 99%

Regenerating Gene Protein as a Novel Autoantigen in the Pathogenesis of Sjögren’s Syndrome

et al. 2015

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Sjögren's syndrome, an autoimmune disease characterized by exocrine gland dysfunction leading to dry mouth and dry eye diseases, is typified by lymphoplasmacytic infiltrations and a progressive destruction of the salivary and lacrimal glands. Despite an ever-increasing focus on identifying the underlying etiology of Sjögren's syndrome, the factors that initiate this autoimmune disease and the mechanisms that cause the subsequent exocrine gland dysfunction remain a mystery. The original explanatory concept for the pathogenesis of Sjögren's syndrome proposed a specific, self-perpetuating, immune-mediated loss of acinar and ductal cells as the principal cause of salivary gland dysfunction. We highlight the possible involvement of regenerating gene (Reg) in the regeneration and destruction of salivary gland acinar and ductal cells in Sjögren's syndrome. The Reg gene was originally isolated as a gene specifically overexpressed in regenerating pancreatic islets and constitutes a growth factor family (Reg family). We describe how salivary gland dysfunction is initiated and maintained and how it can be regenerated or progressed, mediated by the Reg gene, Reg protein, and anti-REG autoantibodies in Sjögren's syndrome. OPEN ACCESSAntibodies 2015, 4 410

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Construction and function of a recombinant adenovirus encoding a human aquaporin 1-green fluorescent protein fusion product

Cited by 15 publications

References 30 publications

Aquaporin-1, Nothing but a Water Channel

Aquaporin-1, Nothing but a Water Channel

Significance of Interleukin-6/STAT Pathway for the Gene Expression of REG Iα, a New Autoantigen in Sjögren’s Syndrome Patients, in Salivary Duct Epithelial Cells

Regenerating Gene Protein as a Novel Autoantigen in the Pathogenesis of Sjögren’s Syndrome

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