17 results on '"Caroline Weydert"'
Search Results
2. Correction to: HIV-1 IN/Pol recruits LEDGF/p75 into viral particles
- Author
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Belete Ayele Desimmie, Caroline Weydert, Rik Schrijvers, Sofie Vets, Jonas Demeulemeester, Paul Proost, Igor Paron, Jan De Rijck, Jan Mast, Norbert Bannert, Rik Gijsbers, Frauke Christ, and Zeger Debyser
- Subjects
Immunologic diseases. Allergy ,RC581-607 - Abstract
An amendment to this paper has been published and can be accessed via the original article.
- Published
- 2020
- Full Text
- View/download PDF
3. Correction to: LEDGINs inhibit late stage HIV-1 replication by modulating integrase multimerization in the virions
- Author
-
Belete Ayele Desimmie, Rik Schrijvers, Jonas Demeulemeester, Doortje Borrenberghs, Caroline Weydert, Wannes Thys, Sofie Vets, Barbara Van Remoortel, Johan Hofkens, Jan De Rijck, Jelle Hendrix, Norbert Bannert, Rik Gijsbers, Frauke Christ, and Zeger Debyser
- Subjects
Immunologic diseases. Allergy ,RC581-607 - Abstract
An amendment to this paper has been published and can be accessed via the original article.
- Published
- 2020
- Full Text
- View/download PDF
4. LEDGIN-mediated Inhibition of Integrase–LEDGF/p75 Interaction Reduces Reactivation of Residual Latent HIV
- Author
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Lenard S. Vranckx, Jonas Demeulemeester, Suha Saleh, Annegret Boll, Gerlinde Vansant, Rik Schrijvers, Caroline Weydert, Emilie Battivelli, Eric Verdin, Anna Cereseto, Frauke Christ, Rik Gijsbers, and Zeger Debyser
- Subjects
HIV latency ,HIV remission ,Integration ,LEDGIN ,LEDGF/p75 ,Medicine ,Medicine (General) ,R5-920 - Abstract
Persistence of latent, replication-competent Human Immunodeficiency Virus type 1 (HIV-1) provirus is the main impediment towards a cure for HIV/AIDS (Acquired Immune Deficiency Syndrome). Therefore, different therapeutic strategies to eliminate the viral reservoirs are currently being explored. We here propose a novel strategy to reduce the replicating HIV reservoir during primary HIV infection by means of drug-induced retargeting of HIV integration. A novel class of integration inhibitors, referred to as LEDGINs, inhibit the interaction between HIV integrase and the LEDGF/p75 host cofactor, the main determinant of lentiviral integration site selection. We show for the first time that LEDGF/p75 depletion hampers HIV-1 reactivation in cell culture. Next we demonstrate that LEDGINs relocate and retarget HIV integration resulting in a HIV reservoir that is refractory to reactivation by different latency-reversing agents. Taken together, these results support the potential of integrase inhibitors that modulate integration site targeting to reduce the likeliness of viral rebound.
- Published
- 2016
- Full Text
- View/download PDF
5. Y-box-binding protein 1 supports the early and late steps of HIV replication.
- Author
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Caroline Weydert, Bart van Heertum, Lieve Dirix, Stéphanie De Houwer, Flore De Wit, Jan Mast, Steven J Husson, Katrien Busschots, Renate König, Rik Gijsbers, Jan De Rijck, and Zeger Debyser
- Subjects
Medicine ,Science - Abstract
The human immunodeficiency virus (HIV) depends on cellular proteins, so-called cofactors, to complete its replication cycle. In search for new therapeutic targets we identified the DNA and RNA binding protein Y-box-binding Protein 1 (YB-1) as a cofactor supporting early and late steps of HIV replication. YB-1 depletion resulted in a 10-fold decrease in HIV-1 replication in different cell lines. Dissection of the replication defects revealed that knockdown of YB-1 is associated with a 2- to 5-fold decrease in virion production due to interference with the viral RNA metabolism. Using single-round virus infection experiments we demonstrated that early HIV-1 replication also depends on the cellular YB-1 levels. More precisely, using quantitative PCR and an in vivo nuclear import assay with fluorescently labeled viral particles, we showed that YB-1 knockdown leads to a block between reverse transcription and nuclear import of HIV-1. Interaction studies revealed that YB-1 associates with integrase, although a direct interaction with HIV integrase could not be unambiguously proven. In conclusion, our results indicate that YB-1 affects multiple stages of HIV replication. Future research on the interaction between YB-1 and the virus will reveal whether this protein qualifies as a new antiviral target.
- Published
- 2018
- Full Text
- View/download PDF
6. Fundamental insights into autosomal dominant polycystic kidney disease from human-based cell models
- Author
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Caroline Weydert, Humbert De Smedt, Rudi Vennekens, Peter Janssens, Djalila Mekahli, Jean-Paul Decuypere, Clinical sciences, Faculty of Medicine and Pharmacy, Internal Medicine Specializations, and Nephrology
- Subjects
Nephrology ,medicine.medical_specialty ,TRPP Cation Channels ,medicine.medical_treatment ,Primary Cell Culture ,Cell ,030232 urology & nephrology ,Autosomal dominant polycystic kidney disease ,Computational biology ,Disease ,030204 cardiovascular system & hematology ,Kidney ,Cell Line ,03 medical and health sciences ,0302 clinical medicine ,Human cell lines ,cAMP ,Internal medicine ,medicine ,Humans ,Calcium Signaling ,Cilia ,ADPKD ,Cell Proliferation ,Clinical Trials as Topic ,urogenital system ,Mechanism (biology) ,business.industry ,TOR Serine-Threonine Kinases ,Polycystic Kidney, Autosomal Dominant ,medicine.disease ,Nephrectomy ,Treatment Outcome ,medicine.anatomical_structure ,Mutation ,Tolvaptan ,Pediatrics, Perinatology and Child Health ,mTOR ,Calcium ,Stem cell ,business ,Antidiuretic Hormone Receptor Antagonists ,Glucosidases - Abstract
Several animal- and human-derived models are used in autosomal dominant polycystic kidney disease (ADPKD) research to gain insight in the disease mechanism. However, a consistent correlation between animal and human ADPKD models is lacking. Therefore, established human-derived models are relevant to affirm research results and translate findings into a clinical set-up. In this review, we give an extensive overview of the existing human-based cell models. We discuss their source (urine, nephrectomy and stem cell), immortalisation procedures, genetic engineering, kidney segmental origin and characterisation with nephron segment markers. We summarise the most studied pathways and lessons learned from these different ADPKD models. Finally, we issue recommendations for the derivation of human-derived cell lines and for experimental set-ups with these cell lines. ispartof: PEDIATRIC NEPHROLOGY vol:34 issue:10 pages:1697-1715 ispartof: location:Germany status: published
- Published
- 2018
7. Expanding the role of vasopressin antagonism in polycystic kidney diseases: From adults to children?
- Author
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Stéphanie De Rechter, Peter Janssens, Max C. Liebau, Djalila Mekahli, Caroline Weydert, Karl Martin Wissing, Clinical sciences, Internal Medicine Specializations, Faculty of Medicine and Pharmacy, and Basic (bio-) Medical Sciences
- Subjects
Adult ,0301 basic medicine ,Nephrology ,medicine.medical_specialty ,Vasopressins ,Polycystic Kidney, Autosomal Dominant/drug therapy ,030232 urology & nephrology ,Tolvaptan ,Autosomal dominant polycystic kidney disease ,Renal function ,Kidney ,urologic and male genital diseases ,03 medical and health sciences ,0302 clinical medicine ,Signal Transduction/drug effects ,Arginine vasopressin receptor 2 ,Internal medicine ,Cyclic AMP ,medicine ,Polycystic kidney disease ,Animals ,Cyclic AMP/metabolism ,Humans ,Child ,Vasopressins/metabolism ,urogenital system ,business.industry ,Antidiuretic Hormone Receptor Antagonists/therapeutic use ,Kidney metabolism ,Polycystic Kidney, Autosomal Dominant ,medicine.disease ,Tolvaptan/therapeutic use ,female genital diseases and pregnancy complications ,Autosomal Recessive Polycystic Kidney Disease ,Kidney/metabolism ,030104 developmental biology ,Endocrinology ,Pediatrics, Perinatology and Child Health ,business ,Antidiuretic Hormone Receptor Antagonists ,Signal Transduction ,medicine.drug - Abstract
Polycystic kidney disease (PKD) encompasses a group of genetic disorders that are common causes of renal failure. The two classic forms of PKD are autosomal recessive polycystic kidney disease (ARPKD) and autosomal dominant polycystic kidney disease (ADPKD). Despite their clinical differences, ARPKD and ADPKD share many similarities. Altered intracellular Ca2+ and increased cyclic adenosine monophosphate (cAMP) concentrations have repetitively been described as central anomalies that may alter signaling pathways leading to cyst formation. The vasopressin V2 receptor (V2R) antagonist tolvaptan lowers cAMP in cystic tissues and slows renal cystic progression and kidney function decline when given over 3 years in adult ADPKD patients. Tolvaptan is currently approved for the treatment of rapidly progressive disease in adult ADPKD patients. On the occasion of the recent initiation of a clinical trial with tolvaptan in pediatric ADPKD patients, we aim to describe the most important aspects in the literature regarding the AVP-cAMP axis and the clinical use of tolvaptan in PKD.
- Published
- 2017
8. Correction to: HIV-1 IN/Pol recruits LEDGF/p75 into viral particles
- Author
-
Igor Paron, Frauke Christ, Paul Proost, Norbert Bannert, Zeger Debyser, Jan De Rijck, Jonas Demeulemeester, Belete Ayele Desimmie, Rik Schrijvers, Sofie Vets, Jan Mast, Rik Gijsbers, and Caroline Weydert
- Subjects
lcsh:Immunologic diseases. Allergy ,Infectious Diseases ,biology ,business.industry ,Virology ,Human immunodeficiency virus (HIV) ,medicine ,biology.protein ,Antibody ,medicine.disease_cause ,business ,lcsh:RC581-607 - Abstract
An amendment to this paper has been published and can be accessed via the original article.
- Published
- 2020
9. Correction to: LEDGINs inhibit late stage HIV-1 replication by modulating integrase multimerization in the virions
- Author
-
Rik Gijsbers, Jan De Rijck, Sofie Vets, Jelle Hendrix, Belete Ayele Desimmie, Caroline Weydert, Barbara Van Remoortel, Doortje Borrenberghs, Rik Schrijvers, Zeger Debyser, Norbert Bannert, Wannes Thys, Frauke Christ, Jonas Demeulemeester, and Johan Hofkens
- Subjects
LEDGINs ,lcsh:Immunologic diseases. Allergy ,biology ,viruses ,Research ,Human immunodeficiency virus (HIV) ,Late stage ,Antivirals ,Integrase ,medicine.disease_cause ,Virology ,HIV replication ,Infectious Diseases ,Replication (statistics) ,biology.protein ,medicine ,Integrase multimerization ,Antibody ,lcsh:RC581-607 - Abstract
Background LEDGINs are novel allosteric HIV integrase (IN) inhibitors that target the lens epithelium-derived growth factor (LEDGF)/p75 binding pocket of IN. They block HIV-1 integration by abrogating the interaction between LEDGF/p75 and IN as well as by allosterically inhibiting the catalytic activity of IN. Results Here we demonstrate that LEDGINs reduce the replication capacity of HIV particles produced in their presence. We systematically studied the molecular basis of this late effect of LEDGINs and demonstrate that HIV virions produced in their presence display a severe replication defect. Both the late effect and the previously described, early effect on integration contribute to LEDGIN antiviral activity as shown by time-of-addition, qPCR and infectivity assays. The late effect phenotype requires binding of LEDGINs to integrase without influencing proteolytic cleavage or production of viral particles. LEDGINs augment IN multimerization during virion assembly or in the released viral particles and severely hamper the infectivity of progeny virions. About 70% of the particles produced in LEDGIN-treated cells do not form a core or display aberrant empty cores with a mislocalized electron-dense ribonucleoprotein. The LEDGIN-treated virus displays defective reverse transcription and nuclear import steps in the target cells. The LEDGIN effect is possibly exerted at the level of the Pol precursor polyprotein. Conclusion Our results suggest that LEDGINs modulate IN multimerization in progeny virions and impair the formation of regular cores during the maturation step, resulting in a decreased infectivity of the viral particles in the target cells. LEDGINs thus profile as unique antivirals with combined early (integration) and late (IN assembly) effects on the HIV replication cycle.
- Published
- 2020
10. Lessons Learned: HIV Points the Way Towards Precision Treatment of Mixed-Lineage Leukemia
- Author
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Jan De Rijck, Frauke Christ, Zeger Debyser, Katerina Cermakova, and Caroline Weydert
- Subjects
Models, Molecular ,0301 basic medicine ,HIV Infections ,Computational biology ,Biology ,Toxicology ,Protein–protein interaction ,Small Molecule Libraries ,03 medical and health sciences ,0302 clinical medicine ,medicine ,Animals ,Humans ,Protein Interaction Domains and Motifs ,Molecular Targeted Therapy ,Precision Medicine ,Transcription factor ,Adaptor Proteins, Signal Transducing ,Pharmacology ,Genetics ,Acute leukemia ,Leukemia ,Contraindications ,Histone-Lysine N-Methyltransferase ,Precision medicine ,medicine.disease ,Chromatin ,030104 developmental biology ,030220 oncology & carcinogenesis ,Myeloid-Lymphoid Leukemia Protein ,Transcription Factors - Abstract
Protein-protein interactions are involved in most if not all pathogenic and pathophysiological processes and represent attractive therapeutic targets. Extensive biological and clinical research efforts have led to the identification and validation of several cellular hubs that are crucially involved in disease pathogenesis. An interesting example of such a hub is the lens epithelium-derived growth factor (LEDGF/p75), a protein that tethers multiple unrelated proteins and protein complexes to the chromatin. Its chromatin-tethering ability is linked to at least two unrelated diseases-HIV infection and MLL-rearranged acute leukemia. In this review we discuss recent progress in our understanding of the interaction of LEDGF/p75 with its binding partners and focus on the first steps towards therapies targeting protein-protein interactions of LEDGF/p75.
- Published
- 2016
11. Y-box-binding protein 1 supports the early and late steps of HIV replication
- Author
-
Jan De Rijck, Flore De Wit, Renate König, Lieve Dirix, Jan Mast, Caroline Weydert, Rik Gijsbers, Steven J. Husson, Katrien Busschots, Stéphanie De Houwer, Bart van Heertum, and Zeger Debyser
- Subjects
0301 basic medicine ,RNA viruses ,Small interfering RNA ,Time Factors ,lcsh:Medicine ,RNA-binding protein ,HIV Integrase ,Pathology and Laboratory Medicine ,Virus Replication ,Biochemistry ,chemistry.chemical_compound ,Immunodeficiency Viruses ,STAGE EMBRYONIC-DEVELOPMENT ,Medicine and Health Sciences ,Small interfering RNAs ,lcsh:Science ,IN-VIVO ,GENE-EXPRESSION ,Multidisciplinary ,NUCLEAR EXPRESSION ,VIRAL-RNA ,Cell biology ,Integrase ,Nucleic acids ,Multidisciplinary Sciences ,Medical Microbiology ,Cell Processes ,Viral Pathogens ,Viruses ,HUMAN-IMMUNODEFICIENCY-VIRUS ,293T cells ,Cell lines ,Science & Technology - Other Topics ,RNA, Viral ,Pathogens ,Biological cultures ,MESSENGER-RNA ,Engineering sciences. Technology ,Research Article ,Yellow Fluorescent Protein ,Active Transport, Cell Nucleus ,Biology ,Transfection ,Research and Analysis Methods ,Microbiology ,Virus ,03 medical and health sciences ,Virology ,Retroviruses ,Genetics ,Humans ,Nuclear Import ,Molecular Biology Techniques ,Non-coding RNA ,Microbial Pathogens ,Molecular Biology ,Science & Technology ,Biology and life sciences ,HUMAN-CELLS ,Lentivirus ,lcsh:R ,Organisms ,HIV ,Proteins ,Correction ,Cell Biology ,Reverse Transcription ,Y box binding protein 1 ,Reverse transcriptase ,Viral Replication ,Gene regulation ,Luminescent Proteins ,030104 developmental biology ,Viral replication ,chemistry ,TRANSCRIPTION START SITES ,biology.protein ,HIV-1 ,RNA ,lcsh:Q ,Gene expression ,Y-Box-Binding Protein 1 ,HOST PROTEINS ,DNA ,HeLa Cells - Abstract
The human immunodeficiency virus (HIV) depends on cellular proteins, so-called cofactors, to complete its replication cycle. In search for new therapeutic targets we identified the DNA and RNA binding protein Y-box-binding Protein 1 (YB-1) as a cofactor supporting early and late steps of HIV replication. YB-1 depletion resulted in a 10-fold decrease in HIV-1 replication in different cell lines. Dissection of the replication defects revealed that knockdown of YB-1 is associated with a 2- to 5-fold decrease in virion production due to interference with the viral RNA metabolism. Using single-round virus infection experiments we demonstrated that early HIV-1 replication also depends on the cellular YB-1 levels. More precisely, using quantitative PCR and an in vivo nuclear import assay with fluorescently labeled viral particles, we showed that YB-1 knockdown leads to a block between reverse transcription and nuclear import of HIV-1. Interaction studies revealed that YB-1 associates with integrase, although a direct interaction with HIV integrase could not be unambiguously proven. In conclusion, our results indicate that YB-1 affects multiple stages of HIV replication. Future research on the interaction between YB-1 and the virus will reveal whether this protein qualifies as a new antiviral target. ispartof: PLOS ONE vol:13 issue:7 ispartof: location:United States status: published
- Published
- 2017
12. LEDGIN-mediated Inhibition of Integrase–LEDGF/p75 Interaction Reduces Reactivation of Residual Latent HIV
- Author
-
Gerlinde Vansant, Emilie Battivelli, Frauke Christ, Annegret Boll, Zeger Debyser, Rik Gijsbers, Lenard Vranckx, Anna Cereseto, Rik Schrijvers, Jonas Demeulemeester, Suha Saleh, Eric Verdin, and Caroline Weydert
- Subjects
CD4-Positive T-Lymphocytes ,0301 basic medicine ,HIV integration ,Integration ,lcsh:Medicine ,Integrase inhibitor ,HIV Infections ,HIV Integrase ,Virus Replication ,LEDGF/p75 ,Virus latency ,LEDGIN ,lcsh:R5-920 ,biology ,Adaptor Proteins ,virus diseases ,General Medicine ,Provirus ,Virus Latency ,3. Good health ,Integrase ,Protein Transport ,Infectious Diseases ,HIV remission ,5.1 Pharmaceuticals ,Public Health and Health Services ,HIV/AIDS ,Development of treatments and therapeutic interventions ,lcsh:Medicine (General) ,HIV latency ,Infection ,Transcription ,Research Paper ,Protein Binding ,Virus Integration ,Clinical Sciences ,General Biochemistry, Genetics and Molecular Biology ,Cell Line ,03 medical and health sciences ,Medicine, General & Internal ,Genetic ,Acquired immunodeficiency syndrome (AIDS) ,medicine ,Humans ,HIV Integrase Inhibitors ,Biology ,Cell Nucleus ,lcsh:R ,Signal Transducing ,medicine.disease ,Virology ,Good Health and Well Being ,030104 developmental biology ,Viral replication ,Immunology ,HIV-1 ,biology.protein ,Virus Activation ,Transcription Factors - Abstract
Persistence of latent, replication-competent Human Immunodeficiency Virus type 1 (HIV-1) provirus is the main impediment towards a cure for HIV/AIDS (Acquired Immune Deficiency Syndrome). Therefore, different therapeutic strategies to eliminate the viral reservoirs are currently being explored. We here propose a novel strategy to reduce the replicating HIV reservoir during primary HIV infection by means of drug-induced retargeting of HIV integration. A novel class of integration inhibitors, referred to as LEDGINs, inhibit the interaction between HIV integrase and the LEDGF/p75 host cofactor, the main determinant of lentiviral integration site selection. We show for the first time that LEDGF/p75 depletion hampers HIV-1 reactivation in cell culture. Next we demonstrate that LEDGINs relocate and retarget HIV integration resulting in a HIV reservoir that is refractory to reactivation by different latency-reversing agents. Taken together, these results support the potential of integrase inhibitors that modulate integration site targeting to reduce the likeliness of viral rebound., Highlights • LEDGF/p75 depletion hampers HIV reactivation in cell culture. • LEDGINs relocate and retarget authentic HIV integration. • LEDGIN treatment results in quiescent residual HIV provirus which is less susceptible to reactivation. • LEDGIN treatment during primary HIV infection may lead to an HIV remission. Different strategies to cure HIV infection are being explored. Although complete eradication of the HIV provirus is the ultimate goal, disease remission allowing treatment interruption without viral rebound would constitute a significant leap forward. HIV integration site selection is orchestrated by LEDGF/p75. The advent of LEDGINs, that block the interaction between integrase and LEDGF/p75, allowed us to examine the hypothesis that interference with HIV integration site selection would yield integration sites that are less optimal for productive infection. Here we provide evidence in cell culture that LEDGIN treatment during acute HIV infection yields an HIV reservoir refractory to reactivation.
- Published
- 2017
13. HIV Virions as Nanoscopic Test Tubes for Probing Oligomerization of the Integrase Enzyme
- Author
-
Wannes Thys, Peter Dedecker, Jonas Demeulemeester, Doortje Borrenberghs, Zeger Debyser, Johan Hofkens, Jelle Hendrix, Caroline Weydert, and Susana Rocha
- Subjects
Models, Molecular ,single-molecule fluorescence microscopy ,viruses ,General Physics and Astronomy ,HIV Integrase ,nanoscopy ,Article ,Protein–protein interaction ,HIV-1 integrase ,Fluorescence Resonance Energy Transfer ,Humans ,Nanotechnology ,General Materials Science ,Protein Structure, Quaternary ,Nanoscopic scale ,chemistry.chemical_classification ,biology ,HEK 293 cells ,Virion ,General Engineering ,Fluorescence ,Virology ,In vitro ,stoichiometry ,Integrase ,HEK293 Cells ,Förster resonance energy transfer ,Enzyme ,chemistry ,Biocatalysis ,HIV-1 ,Biophysics ,biology.protein ,protein−protein interactions ,Protein Multimerization - Abstract
Employing viruses as nanoscopic lipid-enveloped test tubes allows the miniaturization of protein–protein interaction (PPI) assays while preserving the physiological environment necessary for particular biological processes. Applied to the study of the human immunodeficiency virus type 1 (HIV-1), viral biology and pathology can also be investigated in novel ways, both in vitro as well as in infected cells. In this work we report on an experimental strategy that makes use of engineered HIV-1 viral particles, to allow for probing PPIs of the HIV-1 integrase (IN) inside viruses with single-molecule Förster resonance energy transfer (FRET) using fluorescent proteins (FP). We show that infectious fluorescently labeled viruses can be obtained and that the quantity of labels can be accurately measured and controlled inside individual viral particles. We demonstrate, with proper control experiments, the formation of IN oligomers in single viral particles and inside viral complexes in infected cells. Finally, we show a clear effect on IN oligomerization of small molecule inhibitors of interactions of IN with its natural human cofactor LEDGF/p75, corroborating that IN oligomer enhancing drugs are active already at the level of the virus and strongly suggesting the presence of a dynamic, enhanceable equilibrium between the IN dimer and tetramer in viral particles. Although applied to the HIV-1 IN enzyme, our methodology for utilizing HIV virions as nanoscopic test tubes for probing PPIs is generic, i.e., other PPIs targeted into the HIV-1, or PPIs targeted into other viruses, can potentially be studied with a similar strategy.
- Published
- 2014
14. Targeting Virus-host Interactions of HIV Replication
- Author
-
Jan De Rijck, Frauke Christ, Caroline Weydert, and Zeger Debyser
- Subjects
viruses ,Human immunodeficiency virus (HIV) ,Nerve Tissue Proteins ,Plasma protein binding ,Computational biology ,HIV Integrase ,Receptors, Nerve Growth Factor ,medicine.disease_cause ,Virus Replication ,Protein–protein interaction ,chemistry.chemical_compound ,Drug Discovery ,medicine ,Humans ,Maraviroc ,Virus host ,biology ,General Medicine ,Virology ,Replication (computing) ,Integrase ,Viral replication ,chemistry ,biology.protein ,HIV-1 ,Intercellular Signaling Peptides and Proteins ,Protein Binding - Abstract
Cellular proteins that are hijacked by HIV in order to complete its replication cycle, form attractive new targets for antiretroviral therapy. In particular, the protein-protein interactions between these cellular proteins (cofactors) and viral proteins are of great interest to develop new therapies. Research efforts have led to the validation of different cofactors and some successes in therapeutic applications. Maraviroc, the first cofactor inhibitor approved for human medicinal use, provided a proof of concept. Furthermore, compounds developed as Integrase-LEDGF/p75 interaction inhibitors (LEDGINs) have advanced to early clinical trials. Other compounds targeting cofactors and cofactor-viral protein interactions are currently under development. Likewise, interactions between cellular restriction factors and their counteracting HIV protein might serve as interesting targets in order to impair HIV replication. In this respect, compounds targeting the Vif-APOBEC3G interaction have been described. In this review, we focus on compounds targeting the Integrase-LEDGF/p75 interaction, the Tat-P-TEFb interaction and the Vif-APOBEC3G interaction. Additionally we give an overview of currently discovered compounds presumably targeting cellular cofactor-HIV protein interactions. ispartof: Current Topics in Medicinal Chemistry vol:16 issue:10 pages:1167-1190 ispartof: location:United Arab Emirates status: published
- Published
- 2015
15. HIV-1 IN/Pol recruits LEDGF/p75 into viral particles
- Author
-
Jan Mast, Jan De Rijck, Sofie Vets, Frauke Christ, Zeger Debyser, Jonas Demeulemeester, Rik Gijsbers, Rik Schrijvers, Igor Paron, Norbert Bannert, Caroline Weydert, Belete Ayele Desimmie, and Paul Proost
- Subjects
musculoskeletal diseases ,medicine.medical_treatment ,Virus Integration ,viruses ,Assembly ,Integrase inhibitor ,Virus Replication ,Integrase ,Virus ,HIV Protease ,Virology ,LEDGF/p75 ,medicine ,Humans ,Host factor ,Adaptor Proteins, Signal Transducing ,Protease cleavage sites ,Protease ,biology ,Research ,Correction ,biological factors ,Chromatin ,Infectious Diseases ,Viral replication ,nervous system ,pol Gene Products, Human Immunodeficiency Virus ,Host-Pathogen Interactions ,Proteolysis ,biology.protein ,HIV-1 ,sense organs ,Transcription Factors - Abstract
Background The dynamic interaction between HIV and its host governs the replication of the virus and the study of the virus-host interplay is key to understand the viral lifecycle. The host factor lens epithelium-derived growth factor (LEDGF/p75) tethers the HIV preintegration complex to the chromatin through a direct interaction with integrase (IN). Small molecules that bind the LEDGF/p75 binding pocket of the HIV IN dimer (LEDGINs) block HIV replication through a multimodal mechanism impacting early and late stage replication including HIV maturation. Furthermore, LEDGF/p75 has been identified as a Pol interaction partner. This raised the question whether LEDGF/p75 besides acting as a molecular tether in the target cell, also affects late steps of HIV replication. Results LEDGF/p75 is recruited into HIV-1 particles through direct interaction with the viral IN (or Pol polyprotein) and is a substrate for HIV-1 protease. Incubation in the presence of HIV-1 protease inhibitors resulted in detection of full-length LEDGF/p75 in purified viral particles. We also demonstrate that inhibition of LEDGF/p75-IN interaction by specific mutants or LEDGINs precludes incorporation of LEDGF/p75 in virions, underscoring the specificity of the uptake. LEDGF/p75 depletion did however not result in altered LEDGIN potency. Conclusion Together, these results provide evidence for an IN/Pol mediated uptake of LEDGF/p75 in viral particles and a specific cleavage by HIV protease. Understanding of the possible role of LEDGF/p75 or its cleavage fragments in the viral particle awaits further experimentation. Electronic supplementary material The online version of this article (doi:10.1186/s12977-014-0134-4) contains supplementary material, which is available to authorized users.
- Published
- 2015
16. LEDGINs inhibit late stage HIV-1 replication by modulating integrase multimerization in the virions
- Author
-
Jonas Demeulemeester, Belete Ayele Desimmie, Barbara Van Remoortel, Frauke Christ, Norbert Bannert, Caroline Weydert, Doortje Borrenberghs, Jan De Rijck, Wannes Thys, Rik Schrijvers, Zeger Debyser, Johan Hofkens, Jelle Hendrix, Sofie Vets, and Rik Gijsbers
- Subjects
viruses ,HIV Integrase ,Virus Replication ,Virus ,03 medical and health sciences ,Virology ,Humans ,HIV Integrase Inhibitors ,Cells, Cultured ,030304 developmental biology ,Ribonucleoprotein ,Infectivity ,0303 health sciences ,biology ,Virus Assembly ,030302 biochemistry & molecular biology ,Correction ,Reverse transcriptase ,3. Good health ,Integrase ,Infectious Diseases ,Viral replication ,Virion assembly ,biology.protein ,HIV-1 ,Leukocytes, Mononuclear ,Nuclear transport ,Protein Multimerization - Abstract
Background LEDGINs are novel allosteric HIV integrase (IN) inhibitors that target the lens epithelium-derived growth factor (LEDGF)/p75 binding pocket of IN. They block HIV-1 integration by abrogating the interaction between LEDGF/p75 and IN as well as by allosterically inhibiting the catalytic activity of IN. Results Here we demonstrate that LEDGINs reduce the replication capacity of HIV particles produced in their presence. We systematically studied the molecular basis of this late effect of LEDGINs and demonstrate that HIV virions produced in their presence display a severe replication defect. Both the late effect and the previously described, early effect on integration contribute to LEDGIN antiviral activity as shown by time-of-addition, qPCR and infectivity assays. The late effect phenotype requires binding of LEDGINs to integrase without influencing proteolytic cleavage or production of viral particles. LEDGINs augment IN multimerization during virion assembly or in the released viral particles and severely hamper the infectivity of progeny virions. About 70% of the particles produced in LEDGIN-treated cells do not form a core or display aberrant empty cores with a mislocalized electron-dense ribonucleoprotein. The LEDGIN-treated virus displays defective reverse transcription and nuclear import steps in the target cells. The LEDGIN effect is possibly exerted at the level of the Pol precursor polyprotein. Conclusion Our results suggest that LEDGINs modulate IN multimerization in progeny virions and impair the formation of regular cores during the maturation step, resulting in a decreased infectivity of the viral particles in the target cells. LEDGINs thus profile as unique antivirals with combined early (integration) and late (IN assembly) effects on the HIV replication cycle.
- Published
- 2013
17. The binding mechanism of a peptidic cyclic serine protease inhibitor
- Author
-
Masood Hosseini, Knud J. Jensen, Jan K. Jensen, Niels Chr. Nielsen, Hans Peter Sørensen, Anders Malmendal, Peter A. Andreasen, Caroline Weydert, Anni Christensen, Longguang Jiang, Mingdong Huang, Cai Yuan, Anna Sigrid Pii Svane, Zhuo Chen, and Jakob Nielsen
- Subjects
Proteases ,Serine Proteinase Inhibitors ,Stereochemistry ,medicine.medical_treatment ,Molecular Sequence Data ,Peptide ,Peptides, Cyclic ,Serine ,Structural Biology ,medicine ,Humans ,Amino Acid Sequence ,Molecular Biology ,Serine protease ,chemistry.chemical_classification ,Protease ,biology ,Chemistry ,Isothermal titration calorimetry ,Surface Plasmon Resonance ,Protease inhibitor (biology) ,Biochemistry ,biology.protein ,Plasminogen activator ,medicine.drug ,Protein Binding - Abstract
Serine proteases are classical objects for studies of catalytic and inhibitory mechanisms as well as interesting as therapeutic targets. Since small-molecule serine protease inhibitors generally suffer from specificity problems, peptidic inhibitors, isolated from phage-displayed peptide libraries, have attracted considerable attention. Here, we have investigated the mechanism of binding of peptidic inhibitors to serine protease targets. Our model is upain-1 (CSWRGLENHRMC), a disulfide-bond-constrained competitive inhibitor of human urokinase-type plasminogen activator with a noncanonical inhibitory mechanism and an unusually high specificity. Using a number of modified variants of upain-1, we characterised the upain-1–urokinase-type plasminogen activator complex using X-ray crystal structure analysis, determined a model of the peptide in solution by NMR spectroscopy, and analysed binding kinetics and thermodynamics by surface plasmon resonance and isothermal titration calorimetry. We found that upain-1 changes both main-chain conformation and side-chain orientations as it binds to the protease, in particular its Trp3 residue and the surrounding backbone. The properties of upain-1 are strongly influenced by the addition of three to four amino acids long N-terminal and C-terminal extensions to the core, disulfide-bond-constrained sequence: The C-terminal extension stabilises the solution structure compared to the core peptide alone, and the protease-bound structure of the peptide is stabilised by intrapeptide contacts between the N-terminal extension and the core peptide around Trp3. These results provide a uniquely detailed description of the binding of a peptidic protease inhibitor to its target and are of general importance in the development of peptidic inhibitors with high specificity and new inhibitory mechanisms.
- Published
- 2011
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