Abstract
DNA-protein interaction in the 5 upstream polymorphic region of the dopamine D4 receptor (DRD4) gene was analyzed by capillary electrophoretic mobility shift assay (CEMSA). The sequence of interest was amplified using a fluorescent primer and applied as a probe in the binding assays with HeLa nuclear extract. Serial dilution of the probe resulted in a concentration dependent DNA-protein complex formation. Sp 1 specific oligonucleotide competitor significantly inhibited the DNA-protein complex formation. A non-specific competitor, differing only in three base pairs, showed weaker effect pointing to the contribution of the Sp 1 recognition sequence in the complex. Polymorphic competitors were also prepared from homozygous individuals possessing either duplicated (2x120 bp) or single copy (1x120bp) of the 120 bp repeat sequence and were used against the Sp 1 specific probe in competition assays. Our data provide experimental evidence for the binding of Sp 1 to the 120 bp duplicated sequence of the DRD4 5 upstream region and suggest enhanced binding capacity of the duplicated form.
Keywords: capillary electrophoresis, electrophoretic mobility shift assay, dopamine d4 receptor
Current Medicinal Chemistry
Title: Capillary Electrophoresis Study on DNA-Protein Complex Formation in the Polymorphic 5 Upstream Region of the Dopamine D4 Receptor (DRD4) Gene
Volume: 11 Issue: 8
Author(s): Z. Ronai, A. Guttman, G. Keszler and M. Sasvari-Szekely
Affiliation:
Keywords: capillary electrophoresis, electrophoretic mobility shift assay, dopamine d4 receptor
Abstract: DNA-protein interaction in the 5 upstream polymorphic region of the dopamine D4 receptor (DRD4) gene was analyzed by capillary electrophoretic mobility shift assay (CEMSA). The sequence of interest was amplified using a fluorescent primer and applied as a probe in the binding assays with HeLa nuclear extract. Serial dilution of the probe resulted in a concentration dependent DNA-protein complex formation. Sp 1 specific oligonucleotide competitor significantly inhibited the DNA-protein complex formation. A non-specific competitor, differing only in three base pairs, showed weaker effect pointing to the contribution of the Sp 1 recognition sequence in the complex. Polymorphic competitors were also prepared from homozygous individuals possessing either duplicated (2x120 bp) or single copy (1x120bp) of the 120 bp repeat sequence and were used against the Sp 1 specific probe in competition assays. Our data provide experimental evidence for the binding of Sp 1 to the 120 bp duplicated sequence of the DRD4 5 upstream region and suggest enhanced binding capacity of the duplicated form.
Export Options
About this article
Cite this article as:
Ronai Z., Guttman A., Keszler G. and Sasvari-Szekely M., Capillary Electrophoresis Study on DNA-Protein Complex Formation in the Polymorphic 5 Upstream Region of the Dopamine D4 Receptor (DRD4) Gene, Current Medicinal Chemistry 2004; 11 (8) . https://dx.doi.org/10.2174/0929867043455503
DOI https://dx.doi.org/10.2174/0929867043455503 |
Print ISSN 0929-8673 |
Publisher Name Bentham Science Publisher |
Online ISSN 1875-533X |
Call for Papers in Thematic Issues
Advances in Medicinal Chemistry: From Cancer to Chronic Diseases.
The broad spectrum of the issue will provide a comprehensive overview of emerging trends, novel therapeutic interventions, and translational insights that impact modern medicine. The primary focus will be diseases of global concern, including cancer, chronic pain, metabolic disorders, and autoimmune conditions, providing a broad overview of the advancements in ...read more
Approaches to the treatment of chronic inflammation
Chronic inflammation is a hallmark of numerous diseases, significantly impacting global health. Although chronic inflammation is a hot topic, not much has been written about approaches to its treatment. This thematic issue aims to showcase the latest advancements in chronic inflammation treatment and foster discussion on future directions in this ...read more
Cellular and Molecular Mechanisms of Non-Infectious Inflammatory Diseases: Focus on Clinical Implications
The Special Issue covers the results of the studies on cellular and molecular mechanisms of non-infectious inflammatory diseases, in particular, autoimmune rheumatic diseases, atherosclerotic cardiovascular disease and other age-related disorders such as type II diabetes, cancer, neurodegenerative disorders, etc. Review and research articles as well as methodology papers that summarize ...read more
Chalcogen-modified nucleic acid analogues
Chalcogen-modified nucleosides, nucleotides and oligonucleotides have been of great interest to scientific research for many years. The replacement of oxygen in the nucleobase, sugar or phosphate backbone by chalcogen atoms (sulfur, selenium, tellurium) gives these biomolecules unique properties resulting from their altered physical and chemical properties. The continuing interest in ...read more
- Author Guidelines
- Graphical Abstracts
- Fabricating and Stating False Information
- Research Misconduct
- Post Publication Discussions and Corrections
- Publishing Ethics and Rectitude
- Increase Visibility of Your Article
- Archiving Policies
- Peer Review Workflow
- Order Your Article Before Print
- Promote Your Article
- Manuscript Transfer Facility
- Editorial Policies
- Allegations from Whistleblowers
- Announcements
Related Articles
-
Developments of Combretastatin A-4 Derivatives as Anticancer Agents
Current Medicinal Chemistry A New Era of Immunotherapy in Prostate Cancer
Current Molecular Pharmacology Anti-cancer and Anti-angiogenic Properties of Various Natural Pentacyclic Tri-terpenoids and Some of their Chemical Derivatives
Current Organic Chemistry Nanoparticles for Tumor Targeted Therapies and Their Pharmacokinetics
Current Drug Metabolism Combined Treatment with JFKD and Gefitinib Overcomes Drug Resistance in Non-Small Cell Lung Cancer
Current Pharmaceutical Biotechnology Inhibitors of ABL and the ABL-T315I Mutation
Current Topics in Medicinal Chemistry A Preliminary Study for Quantitative Assessment with HFUS (High- Frequency Ultrasound) of Nodular Skin Melanoma Breslow Thickness in Adults Before Surgery: Interdisciplinary Team Experience
Current Radiopharmaceuticals Potential of DNMT and its Epigenetic Regulation for Lung Cancer Therapy
Current Genomics Reduction of Breast Cancer Relapses with Perioperative Non-Steroidal Anti-Inflammatory Drugs: New Findings and a Review
Current Medicinal Chemistry Molecular Docking Studies and Inhibition Properties of Some Antineoplastic Agents against Paraoxonase-I
Anti-Cancer Agents in Medicinal Chemistry Drugging Cell Cycle Kinases in Cancer Therapy
Current Drug Targets Cytotoxic and Chemopreventive Effects of Gemin D Against Different Mutagens Using In Vitro and In Vivo Assays
Anti-Cancer Agents in Medicinal Chemistry Investigation of the Effect of <i>Prunus Amygdalus</i> Amara on the Expression of some Genes of Apoptosis and Immortality in Breast Cancer Cells (MCF- 7)
Current Drug Research Reviews Endothelial Endocytic Pathways: Gates for Vascular Drug Delivery
Current Vascular Pharmacology Turner Syndrome : How Is It Made Up?
Current Genomics Patterns of Self-Medication with Medicinal Plants and Related Adverse Events - A South American Survey
Current Drug Safety Role of Phytochemicals in the Prevention of Menopausal Bone Loss: Evidence from In Vitro and In Vivo, Human Interventional and Pharmacokinetic Studies
Current Medicinal Chemistry Peptides Homing to Tumor Vasculature: Imaging and Therapeutics for Cancer
Recent Patents on Anti-Cancer Drug Discovery Reversal of Tumor Induced Dendritic Cell Paralysis: A Treatment Regimen Against Cancer
Current Immunology Reviews (Discontinued) Urokinase-a Very Popular Cardiovascular Agent
Recent Patents on Cardiovascular Drug Discovery