Evidence map›Paper›PMID 33600497›Full record

ArticlePloS one2021

Characterization of the G protein-coupled receptor kinase 6 promoter reveals a functional CREB binding site.

Maike Stegen, Andrea Engler, Crista Ochsenfarth, Iris Manthey, Jürgen Peters, Winfried Siffert, Ulrich H Frey

Open access · goldAbstract read
In one paragraph

Article in PloS one, 2021. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Cited by 3 papers.

0numbers the graph read from it
0cells of the map it votes in
3citing papers in PubMed
0.2field-weighted citation impact, top 49% of its field
1 · What the graph read from it

What it found

Each row is one number read from the abstract, on the scale the paper reported it, with its interval. Left of the dashed line favours the treatment, right favours the comparator. Under each row is the sentence it came from. New to these charts? A ten-minute tutorial.

The abstract states no effect estimate the extractor could read, or names no intervention and outcome on the map, so this paper lights no cell and moves no belief. It is still indexed, cited and linked below.

2 · The registry

The trial behind it

Trials whose registry record cites this paper, or whose number appears in the abstract. A trial that started after this paper was published is citing it as background, not reporting it.

Neither the registry nor the abstract names a trial number. If this is a trial report, that itself is worth knowing.

3 · Its place in the literature

Who cites it

3 citing papers in PubMed, 3 citations in OpenAlex.

  1. G protein-coupled receptor kinases in hypertension: physiology, pathogenesis, and therapeutic targets.Hypertension research : official journal of the Japanese Society of Hypertension · 2024
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4 · The record

Corrections and comments

PubMed lists nothing against this paper. Absence here is not a guarantee, only a check that was made.

5 · Who and what money

Authors and funding

7 authors at 2 institutions in 1 country.

Maike StegenDepartment of Anaesthesiology and Intensive Care Medicine, Essen University Hospital and University of Duisburg-Essen, Essen, Germany.ORCID 0000-0001-5467-0446
Andrea EnglerDepartment of Anaesthesiology and Intensive Care Medicine, Essen University Hospital and University of Duisburg-Essen, Essen, Germany.
Crista OchsenfarthDepartment of Anaesthesiology, Operative Intensive Care Medicine, Pain and Palliative Medicine, Marien Hospital Herne, Ruhr-University Bochum, Bochum, Germany.ORCID 0000-0002-5744-8661
Iris MantheyInstitute of Pharmacogenetics, Essen University Hospital and University of Duisburg-Essen, Essen, Germany.
Jürgen PetersDepartment of Anaesthesiology and Intensive Care Medicine, Essen University Hospital and University of Duisburg-Essen, Essen, Germany.
Winfried SiffertInstitute of Pharmacogenetics, Essen University Hospital and University of Duisburg-Essen, Essen, Germany.
Ulrich H FreyDepartment of Anaesthesiology and Intensive Care Medicine, Essen University Hospital and University of Duisburg-Essen, Essen, Germany.ORCID 0000-0003-2961-6629
Essen University Hospital · DEUniversitätsklinik Marien Hospital Herne · DE

Funding

No grant is acknowledged in the PubMed record.

6 · The paper itself

Abstract

backgroundG protein-coupled receptor kinase 6 (GRK6) is part of the G protein-coupled receptor kinase family, whose members act as key regulators of seven-transmembrane receptor signalling. GRK6 seems to play a role in regulation of inflammatory processes, but mechanisms of transcriptional regulation of GRK6 expression in inflammatory cell lines have not been characterized. Protein kinase C (PKC) signalling is also involved in inflammatory regulation and an impact of PKC activation on GRK6 protein expression was described previously. Thus, the aim of this study was to 1) characterize the GRK6 promoter, and 2) investigate a potential influence of PKC on GRK6 expression.

methodsFive deletion constructs of the GRK6 promoter were cloned. After transient transfection into a human T cell line, promoter activity was assessed using luciferase reporter gene assays. Putative transcription factor binding sites were identified, mutated, and binding was investigated using electrophoretic mobility shift assays (EMSA). Following stimulation with a PKC activator, GRK6 expression on mRNA and protein levels was assessed by reverse transcriptase qPCR and Western blots.

resultsInvestigation of the GRK6 promoter revealed a putative cAMP responsive element (CRE), whose mutation led to decreased promoter activity (p = 0.0006). Functionality of the CRE binding protein (CREB) binding site was verified in EMSA blots. Stimulation with a PKC activator resulted in decreased GRK6 promoter activity (p = 0.0027), mRNA (p = 0.04) and protein expression.

conclusionWe characterized the human GRK6 promoter and identified promoter activity to be influenced by a CREB binding site. PKC might be one determinant contributing to altered GRK6 expression.

Indexed as

Base SequenceBinding SitesCyclic AMP Response Element-Binding ProteinElectrophoretic Mobility Shift AssayG-Protein-Coupled Receptor KinasesHumansJurkat CellsMutagenesis, Site-DirectedPromoter Regions, GeneticProtein BindingProtein Kinase CResponse ElementsRNA, MessengerCREB1 protein, humanCyclic AMP Response Element-Binding ProteinG-protein-coupled receptor kinase 6G-Protein-Coupled Receptor KinasesProtein Kinase CRNA, Messenger

Identifiers

PMID33600497
PMCPMC7891717
OpenAlexW3129849189

What OpenQuestion holds

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Registered trials

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Read under generation 80e0d062 · epoch 390. Bibliography from PubMed, PubMed Central and OpenAlex; grants from NIH RePORTER; trial links from ClinicalTrials.gov; estimates, votes and beliefs from the OpenQuestion graph.