The Experts below are selected from a list of 2274 Experts worldwide ranked by ideXlab platform
Laurie J Ozelius - One of the best experts on this subject based on the ideXlab platform.
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mutations in THAP1 dyt6 reveal that diverse dystonia genes disrupt similar neuronal pathways and functions
PLOS Genetics, 2018Co-Authors: Zuchra Zakirova, Laurie J Ozelius, Justine Bonet, Tomas Fanutza, Ben Readhead, Weijia Zhang, Genevieve Beauvais, Thomas P. Zwaka, Robert D. Blitzer, Pedro GonzalezalegreAbstract:Dystonia is characterized by involuntary muscle contractions. Its many forms are genetically, phenotypically and etiologically diverse and it is unknown whether their pathogenesis converges on shared pathways. Mutations in THAP1 [THAP (Thanatos-associated protein) domain containing, apoptosis associated protein 1], a ubiquitously expressed transcription factor with DNA binding and protein-interaction domains, cause dystonia, DYT6. There is a unique, neuronal 50-kDa THAP1-like immunoreactive species, and THAP1 levels are auto-regulated on the mRNA level. However, THAP1 downstream targets in neurons, and the mechanism via which it causes dystonia are largely unknown. We used RNA-Seq to assay the in vivo effect of a heterozygote THAP1 C54Y or ΔExon2 allele on the gene transcription signatures in neonatal mouse striatum and cerebellum. Enriched pathways and gene ontology terms include eIF2α Signaling, Mitochondrial Dysfunction, Neuron Projection Development, Axonal Guidance Signaling, and Synaptic LongTerm Depression, which are dysregulated in a genotype and tissue-dependent manner. Electrophysiological and neurite outgrowth assays were consistent with those enrichments, and the plasticity defects were partially corrected by salubrinal. Notably, several of these pathways were recently implicated in other forms of inherited dystonia, including DYT1. We conclude that dysfunction of these pathways may represent a point of convergence in the pathophysiology of several forms of inherited dystonia.
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Mutations in THAP1/DYT6 reveal that diverse dystonia genes disrupt similar neuronal pathways and functions
PLoS genetics, 2018Co-Authors: Zuchra Zakirova, Laurie J Ozelius, Justine Bonet, Tomas Fanutza, Weijia Zhang, Genevieve Beauvais, Thomas P. Zwaka, Benjamin Readhead, Robert D. BlitzerAbstract:Dystonia is characterized by involuntary muscle contractions. Its many forms are genetically, phenotypically and etiologically diverse and it is unknown whether their pathogenesis converges on shared pathways. Mutations in THAP1 [THAP (Thanatos-associated protein) domain containing, apoptosis associated protein 1], a ubiquitously expressed transcription factor with DNA binding and protein-interaction domains, cause dystonia, DYT6. There is a unique, neuronal 50-kDa THAP1-like immunoreactive species, and THAP1 levels are auto-regulated on the mRNA level. However, THAP1 downstream targets in neurons, and the mechanism via which it causes dystonia are largely unknown. We used RNA-Seq to assay the in vivo effect of a heterozygote THAP1 C54Y or ΔExon2 allele on the gene transcription signatures in neonatal mouse striatum and cerebellum. Enriched pathways and gene ontology terms include eIF2α Signaling, Mitochondrial Dysfunction, Neuron Projection Development, Axonal Guidance Signaling, and Synaptic LongTerm Depression, which are dysregulated in a genotype and tissue-dependent manner. Electrophysiological and neurite outgrowth assays were consistent with those enrichments, and the plasticity defects were partially corrected by salubrinal. Notably, several of these pathways were recently implicated in other forms of inherited dystonia, including DYT1. We conclude that dysfunction of these pathways may represent a point of convergence in the pathophysiology of several forms of inherited dystonia.
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Global analysis of differential gene expression in striatum and cerebellum of THAP1+/- or THAP1C54Y/+ vs WT.
2018Co-Authors: Zuchra Zakirova, Laurie J Ozelius, Justine Bonet, Tomas Fanutza, Ben Readhead, Weijia Zhang, Genevieve Beauvais, Thomas P. Zwaka, Robert D. BlitzerAbstract:RNA-Seq was used to identify differentially expressed genes (DEGs) in the heterozygote THAP1+/- and THAP1C54Y P1 striatum and cerebellum as compared to WT. Diagrams show number of total DEGs as well as the number of up- or down-regulated genes in the (A) THAP1+/- striatum vs WT (B) THAP1C54Y striatum vs WT. (C) Venn diagrams show the number of overlapping DEGs (total, up-regulated or down-regulated) between THAP1+/- and THAP1C54Y/+ relative to WT striatum. Diagrams show number of total DEGs as well as the number of up- or down-regulated genes in the (D) THAP1+/- cerebellum vs WT (E) THAP1C54Y/+ cerebellum vs WT. (F) Venn diagrams show the number of overlapping DEGs (total, up-regulated or down-regulated) between THAP1+/- and THAP1C54Y/+ relative to WT cerebellum. Cogged gears in panels A, B, D and E represent the number and the direction of the differentially expressed genes for each genotype and brain region, as follows: up-regulated genes (turquoise color, upward right arrow); down-regulated genes (dark blue color, downward left arrow); total [number] of genes (purple color, downward right arrow).
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Inhibition of eIF2α phosphatase rescues mGluR-LTD, but not synaptically-induced LTD.
2018Co-Authors: Zuchra Zakirova, Laurie J Ozelius, Justine Bonet, Tomas Fanutza, Ben Readhead, Weijia Zhang, Genevieve Beauvais, Thomas P. Zwaka, Robert D. BlitzerAbstract:The summary graphs in the right panels show mean ± SEM for the final 5 min of recording. (A) LTD after treatment with group 1 agonist DHPG (100 μM, applied during the gap in the graph) was reduced in THAP1+/- slices, and pretreatment with Sal003 (20 μM) (eIF2α phosphatase inhibitor) rescued LTD in THAP1+/- slices. (B) In high frequency stimulation (HFS)-induced LTD, Sal003 (10 μM) failed to reverse the deficit observed in THAP1+/- slices. Numbers in parentheses indicate number of slices/number of mice. Representative traces are shown during baseline period (solid lines) and at the end of the recording period (dashed lines). Calibrations: 1 mV / 5 ms. Asterisks indicate p < .05 (*) or p < .01 (**) by ANOVAs followed by Newman-Keuls post-hoc tests. See also S9 Table.
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Top canonical pathways and gene ontology terms enriched in striatum and cerebellum of THAP1+/- and THAP1C54Y/+ relative to WT.
2018Co-Authors: Zuchra Zakirova, Laurie J Ozelius, Justine Bonet, Tomas Fanutza, Ben Readhead, Weijia Zhang, Genevieve Beauvais, Thomas P. Zwaka, Robert D. BlitzerAbstract:(A,B) Top canonical pathways as determined by IPA analysis, and (C,D) DAVID Gene Ontology (GO) terms show an enrichment of biological process based on the list of significant DEGs (DEseq p < 0.05) in the striatum of THAP1+/- and THAP1C54Y/+ relative to WT. (E,F) Top canonical pathways and (G,H) DAVID GO terms based on the list of significant DEGs in the cerebellum of THAP1+/- and THAP1C54Y/+ relative to WT.
Alain Milon - One of the best experts on this subject based on the ideXlab platform.
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Structural determinants of specific DNA-recognition by the THAP zinc finger
Nucleic acids research, 2010Co-Authors: Sébastien Campagne, Olivier Saurel, Virginie Gervais, Alain MilonAbstract:Human THAP1 is the prototype of a large family of cellular factors sharing an original THAP zinc-finger motif responsible for DNA binding. Human THAP1 regulates endothelial cell proliferation and G1/S cell-cycle progression, through modulation of pRb/E2F cell-cycle target genes including rrm1. Recently, mutations in THAP1 have been found to cause DYT6 primary torsion dystonia, a human neurological disease. We report here the first 3D structure of the complex formed by the DNA-binding domain of THAP1 and its specific DNA target (THABS) found within the rrm1 target gene. The THAP zinc finger uses its double-stranded β-sheet to fill the DNA major groove and provides a unique combination of contacts from the β-sheet, the N-terminal tail and surrounding loops toward the five invariant base pairs of the THABS sequence. Our studies reveal unprecedented insights into the specific DNA recognition mechanisms within this large family of proteins controlling cell proliferation, cell cycle and pluripotency.
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Structure-Function Analysis of the THAP Zinc Finger of THAP1, a Large C2CH DNA-binding Module Linked to Rb/E2F Pathways
Journal of Biological Chemistry, 2008Co-Authors: Damien Bessiere, Chrystelle Lacroix, Vincent Ecochard, Sébastien Campagne, Valérie Guillet, Lionel Mourey, Frédéric Lopez, Jerzy Czaplicki, Pascal Demange, Alain MilonAbstract:THAP1, the founding member of a previously uncharacterized large family of cellular proteins (THAP proteins), is a sequence-specific DNA-binding factor that has recently been shown to regulate cell proliferation through modulation of pRb/E2F cell cycle target genes. THAP1 shares its DNA-binding THAP zinc finger domain with Drosophila P element transposase, zebrafish E2F6, and several nematode proteins interacting genetically with the retinoblastoma protein pRb. In this study, we report the three-dimensional structure and structure-function relationships of the THAP zinc finger of human THAP1. Deletion mutagenesis and multidimensional NMR spectroscopy revealed that the THAP domain of THAP1 is an atypical zinc finger of ∼80 residues, distinguished by the presence between the C2CH zinc coordinating residues of a short antiparallel β-sheet interspersed by a long loop-helix-loop insertion. Alanine scanning mutagenesis of this loop-helix-loop motif resulted in the identification of a number of critical residues for DNA recognition. NMR chemical shift perturbation analysis was used to further characterize the residues involved in DNA binding. The combination of the mutagenesis and NMR data allowed the mapping of the DNA binding interface of the THAP zinc finger to a highly positively charged area harboring multiple lysine and arginine residues. Together, these data represent the first structure-function analysis of a functional THAP domain, with demonstrated sequence-specific DNA binding activity. They also provide a structural framework for understanding DNA recognition by this atypical zinc finger, which defines a novel family of cellular factors linked to cell proliferation and pRb/E2F cell cycle pathways in humans, fish, and nematodes.
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structure function analysis of the thap zinc finger of THAP1 a large c2ch dna binding module linked to rb e2f pathways
Journal of Biological Chemistry, 2008Co-Authors: Damien Bessiere, Chrystelle Lacroix, Vincent Ecochard, Sébastien Campagne, Valérie Guillet, Lionel Mourey, Frédéric Lopez, Jerzy Czaplicki, Pascal Demange, Alain MilonAbstract:THAP1, the founding member of a previously uncharacterized large family of cellular proteins (THAP proteins), is a sequence-specific DNA-binding factor that has recently been shown to regulate cell proliferation through modulation of pRb/E2F cell cycle target genes. THAP1 shares its DNA-binding THAP zinc finger domain with Drosophila P element transposase, zebrafish E2F6, and several nematode proteins interacting genetically with the retinoblastoma protein pRb. In this study, we report the three-dimensional structure and structure-function relationships of the THAP zinc finger of human THAP1. Deletion mutagenesis and multidimensional NMR spectroscopy revealed that the THAP domain of THAP1 is an atypical zinc finger of approximately 80 residues, distinguished by the presence between the C2CH zinc coordinating residues of a short antiparallel beta-sheet interspersed by a long loop-helix-loop insertion. Alanine scanning mutagenesis of this loop-helix-loop motif resulted in the identification of a number of critical residues for DNA recognition. NMR chemical shift perturbation analysis was used to further characterize the residues involved in DNA binding. The combination of the mutagenesis and NMR data allowed the mapping of the DNA binding interface of the THAP zinc finger to a highly positively charged area harboring multiple lysine and arginine residues. Together, these data represent the first structure-function analysis of a functional THAP domain, with demonstrated sequence-specific DNA binding activity. They also provide a structural framework for understanding DNA recognition by this atypical zinc finger, which defines a novel family of cellular factors linked to cell proliferation and pRb/E2F cell cycle pathways in humans, fish, and nematodes.
Jianfeng Xiao - One of the best experts on this subject based on the ideXlab platform.
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Clinical and genetic features of cervical dystonia in a large multicenter cohort.
Neurology. Genetics, 2016Co-Authors: Mark S. Ledoux, Jianfeng Xiao, Satya R. Vemula, Joel S. Perlmutter, Misty M. Thompson, Laura J. Wright, H.a. Jinnah, Ami Rosen, Peter Hedera, Cynthia L. ComellaAbstract:Objective: To characterize the clinical and genetic features of cervical dystonia (CD). Methods: Participants enrolled in the Dystonia Coalition biorepository (NCT01373424) with initial manifestation as CD were included in this study (n 5 1,000). Data intake included demographics, family history, and the Global Dystonia Rating Scale. Participants were screened for sequence variants (SVs) in GNAL, THAP1, and Exon 5 of TOR1A. Results: The majority of participants were Caucasian (95%) and female (75%). The mean age at onset and disease duration were 45.5 6 13.6 and 14.6 6 11.8 years, respectively. At the time of assessment, 68.5% had involvement limited to the neck, shoulder(s), and proximal arm(s), whereas 47.4% had dystonia limited to the neck. The remaining 31.5% of the individuals exhibited more extensive anatomical spread. A head tremor was noted in 62% of the patients. Head tremor and laryngeal dystonia were more common in females. Psychiatric comorbidities, mainly depression and anxiety, were reported by 32% of the participants and were more common in females. Family histories of dystonia, parkinsonian disorder, and tremor were present in 14%, 11%, and 29% of the patients, respectively. Pathogenic or likely pathogenic SVs in THAP1, TOR1A ,a ndGNAL were identified in 8 participants (0.8%). Two individuals harbored novel missense SVs in Exon 5 of TOR1A. Synonymous and noncoding SVs in THAP1 and GNAL were identified in 4% of the cohort. Conclusions: Head tremor, laryngeal dystonia, and psychiatric comorbidities are more common in female participants with CD. Coding and noncoding variants in GNAL, THAP1 ,a ndTOR1A make
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Blepharospasm in a multiplex African-American pedigree
Journal of the neurological sciences, 2016Co-Authors: Jianfeng Xiao, Satya R. Vemula, Misty M. Thompson, Mark S. LedouxAbstract:Abstract Background Isolated blepharospasm (BSP) is a late-onset focal dystonia characterized by involuntary contractions of the orbicularis oculi muscles. Genetic studies of BSP have been limited by the paucity of large multiplex pedigrees. Although sequence variants (SVs) in THAP1 have been reported in rare cases of BSP, the genetic causes of this focal dystonia remain largely unknown. Moreover, in the absence of family history and strong in silico or in vitro evidence of deleteriousness, the pathogenicity of novel SVs in THAP1 and other dystonia-associated genes can be indeterminate. Methods A large African-American pedigree with BSP was phenotypically characterized and screened for mutations in THAP1, TOR1A and GNAL with Sanger sequencing. Whole-exome sequencing of the proband was used to examine other dystonia-associated genes for potentially pathogenic SVs. In silico and co-segregation analyses were performed for a novel THAP1 SV identified in the proband. Results Seven family members exhibited increased blinking and/or stereotyped bilateral and synchronous orbicularis oculi spasms with age of onset ranging from early childhood to late adult life (7 to 54 years). The proband was found to harbor a novel THAP1 SV (c.314T > C, p.L105S). However, the p.L105S SV did not co-segregate with blepharospasm in the pedigree. Moreover, in silico analyses suggest that p.L105S is benign. No pathogenic or likely pathogenic SVs in other dystonia-associated genes were identified with whole-exome sequencing. Conclusions Blepharospasm can be familial and may be hereditary in African-Americans. A comprehensive array of in silico tools, and, if possible, co-segregation analysis should be used to classify SVs in dystonia-associated genes.
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A rare sequence variant in intron 1 of THAP1 is associated with primary dystonia
Molecular genetics & genomic medicine, 2014Co-Authors: Satya R. Vemula, Yu Zhao, Jianfeng Xiao, Robert W. Bastian, Zbigniew K. Wszolek, Joel S. Perlmutter, Brad A. Racette, Randal C. Paniello, Ryan J. Uitti, Jay A. Van GerpenAbstract:Although coding variants in THAP1 have been causally associated with primary dystonia, the contribution of noncoding variants remains uncertain. Herein, we examine a previously identified Intron 1 variant (c.71+9C>A, rs200209986). Among 1672 subjects with mainly adult-onset primary dystonia, 12 harbored the variant in contrast to 1/1574 controls (P A variant was associated with an elevated ratio of Isoform 1 (NM_018105) to Isoform 2 (NM_199003) in leukocytes. In silico and minigene analyses indicated that c.71+9C>A alters THAP1 splicing. Lymphoblastoid cells harboring the c.71+9C>A variant showed extensive apoptosis with relatively fewer cells in the G2 phase of the cell cycle. Differentially expressed genes from lymphoblastoid cells revealed that the c.71+9C>A variant exerts effects on DNA synthesis, cell growth and proliferation, cell survival, and cytotoxicity. In aggregate, these data indicate that THAP1 c.71+9C>A is a risk factor for adult-onset primary dystonia.
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Neural expression of the transcription factor THAP1 during development in rat
Neuroscience, 2012Co-Authors: Yu Zhao, Jianfeng Xiao, Suzhen Gong, Joseph A. Clara, Mark S. LedouxAbstract:Abstract Loss of function mutations in THAP1 has been associated with primary generalized and focal dystonia in children and adults. THAP1 encodes a transcription factor (THAP1) that harbors an atypical zinc finger domain and plays a critical role in G 1 -S cell cycle control. Current thinking suggests that dystonia may be a neurodevelopmental circuit disorder. Hence, THAP1 may participate in the development of the nervous system. Herein, we report the neurodevelopmental expression patterns of THAP1 transcript and THAP1 protein from the early postnatal period through adulthood in the rat brain, spinal cord and dorsal root ganglia (DRG). We detected THAP1 transcript and THAP1-immunoreactivity (IR) in the cerebral cortex, cerebellum, striatum, substantia nigra, thalamus, spinal cord and DRG. THAP1 transcript expression was higher in the brain than in spinal cord and DRG at P1 and P7 and declined to similar levels at P14 and later time points in all regions except the cerebellum, where it remained high through adulthood. In the brain, THAP1 expression was highest in early development, particularly in the cerebellum at P7. In addition to Purkinje cells in the cerebellum, THAP1-IR was also localized to pyramidal neurons in the cerebral cortex, relay neurons in the thalamus, medium spiny and cholinergic neurons in the striatum, dopaminergic neurons in the substantia nigra, and pyramidal and interneurons in the hippocampus. In the cerebellar cortex, THAP1-IR was prominently distributed in the perikarya and proximal dendrites of Purkinje cells at early time-points. In contrast, it was more diffusely distributed throughout the dendritic arbor of adult Purkinje cells producing a moderate diffuse staining pattern in the molecular layer. At all time points, nuclear IR was weaker than cytoplasmic IR. The prominent cytoplasmic and developmentally regulated expression of THAP1 suggests that THAP1 may function as part of a cell surface-nucleus signaling cascade involved in terminal neural differentiation.
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Genotype-phenotype correlations in THAP1 dystonia: molecular foundations and description of new cases
Parkinsonism & related disorders, 2012Co-Authors: Mark S. Ledoux, Jianfeng Xiao, Monika Rudzińska, Robert W. Bastian, Zbigniew K. Wszolek, Jay A. Van Gerpen, Andreas Puschmann, Dragana Momčilović, Satya R. Vemula, Yu ZhaoAbstract:An extensive variety of THAP1 sequence variants have been associated with focal, segmental and generalized dystonia with age of onset ranging from 3 to over 60 years. In previous work, we screened 1114 subjects with mainly adult-onset primary dystonia (Neurology 2010; 74:229-238) and identified 6 missense mutations in THAP1. For this report, we screened 750 additional subjects for mutations in coding regions of THAP1 and interrogated all published descriptions of THAP1 phenotypes (gender, age of onset, anatomical distribution of dystonia, family history and site of onset) to explore the possibility of THAP1 genotype-phenotype correlations and facilitate a deeper understanding of THAP1 pathobiology. We identified 5 additional missense mutations in THAP1 (p.A7D, p.K16E, p.S21C, p.R29Q, and p.I80V). Three of these variants are associated with appendicular tremors, which were an isolated or presenting sign in some of the affected subjects. Abductor laryngeal dystonia and mild blepharospasm can be manifestations of THAP1 mutations in some individuals. Overall, mean age of onset for THAP1 dystonia is 16.8 years and the most common sites of onset are the arm and neck, and the most frequently affected anatomical site is the neck. In addition, over half of patients exhibit either cranial or laryngeal involvement. Protein truncating mutations and missense mutations within the THAP domain of THAP1 tend to manifest at an earlier age and exhibit more extensive anatomical distributions than mutations localized to other regions of THAP1.
Chrystelle Lacroix - One of the best experts on this subject based on the ideXlab platform.
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the thap zinc finger protein THAP1 associates with coactivator hcf 1 and o glcnac transferase a link between dyt6 and dyt3 dystonias
Journal of Biological Chemistry, 2010Co-Authors: Raoul Mazars, Chrystelle Lacroix, Corinne Cayrol, Anne-claire Lavigne, Nathalie Ortega, Violette Gautier, Gaelle Huet, Anne Gonzalezdeperedo, Jodi L Vogel, Bernard MonsarratAbstract:THAP1 is a sequence-specific DNA binding factor that regulates cell proliferation through modulation of target genes such as the cell cycle-specific gene RRM1. Mutations in the THAP1 DNA binding domain, an atypical zinc finger (THAP-zf), have recently been found to cause DYT6 dystonia, a neurological disease characterized by twisting movements and abnormal postures. In this study, we report that THAP1 shares sequence characteristics, in vivo expression patterns and protein partners with THAP3, another THAP-zf protein. Proteomic analyses identified HCF-1, a potent transcriptional coactivator and cell cycle regulator, and O-GlcNAc transferase (OGT), the enzyme that catalyzes the addition of O-GlcNAc, as major cellular partners of THAP3. THAP3 interacts with HCF-1 through a consensus HCF-1-binding motif (HBM), a motif that is also present in THAP1. Accordingly, THAP1 was found to bind HCF-1 in vitro and to associate with HCF-1 and OGT in vivo. THAP1 and THAP3 belong to a large family of HCF-1 binding factors since seven other members of the human THAP-zf protein family were identified, which harbor evolutionary conserved HBMs and bind to HCF-1. Chromatin immunoprecipitation (ChIP) assays and RNA interference experiments showed that endogenous THAP1 mediates the recruitment of HCF-1 to the RRM1 promoter during endothelial cell proliferation and that HCF-1 is essential for transcriptional activation of RRM1. Together, our findings suggest HCF-1 is an important cofactor for THAP1. Interestingly, our results also provide an unexpected link between DYT6 and DYT3 (X-linked dystonia-parkinsonism) dystonias because the gene encoding the THAP1/DYT6 protein partner OGT maps within the DYT3 critical region on Xq13.1.
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The THAP-Zinc Finger Protein THAP1 Associates with Coactivator HCF-1 andO-GlcNAc Transferase
Journal of Biological Chemistry, 2010Co-Authors: Raoul Mazars, Chrystelle Lacroix, Corinne Cayrol, Anne Gonzalez-de-peredo, Anne-claire Lavigne, Jodi Vogel, Nathalie Ortega, Violette Gautier, Gaelle Huet, Aurélie RayAbstract:THAP1 is a sequence-specific DNA binding factor that regulates cell proliferation through modulation of target genes such as the cell cycle-specific gene RRM1. Mutations in the THAP1 DNA binding domain, an atypical zinc finger (THAP-zf), have recently been found to cause DYT6 dystonia, a neurological disease characterized by twisting movements and abnormal postures. In this study, we report that THAP1 shares sequence characteristics, in vivo expression patterns and protein partners with THAP3, another THAP-zf protein. Proteomic analyses identified HCF-1, a potent transcriptional coactivator and cell cycle regulator, and O-GlcNAc transferase (OGT), the enzyme that catalyzes the addition of O-GlcNAc, as major cellular partners of THAP3. THAP3 interacts with HCF-1 through a consensus HCF-1-binding motif (HBM), a motif that is also present in THAP1. Accordingly, THAP1 was found to bind HCF-1 in vitro and to associate with HCF-1 and OGT in vivo. THAP1 and THAP3 belong to a large family of HCF-1 binding factors since seven other members of the human THAP-zf protein family were identified, which harbor evolutionary conserved HBMs and bind to HCF-1. Chromatin immunoprecipitation (ChIP) assays and RNA interference experiments showed that endogenous THAP1 mediates the recruitment of HCF-1 to the RRM1 promoter during endothelial cell proliferation and that HCF-1 is essential for transcriptional activation of RRM1. Together, our findings suggest HCF-1 is an important cofactor for THAP1. Interestingly, our results also provide an unexpected link between DYT6 and DYT3 (X-linked dystonia-parkinsonism) dystonias because the gene encoding the THAP1/DYT6 protein partner OGT maps within the DYT3 critical region on Xq13.1.
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Structure-Function Analysis of the THAP Zinc Finger of THAP1, a Large C2CH DNA-binding Module Linked to Rb/E2F Pathways
Journal of Biological Chemistry, 2008Co-Authors: Damien Bessiere, Chrystelle Lacroix, Vincent Ecochard, Sébastien Campagne, Valérie Guillet, Lionel Mourey, Frédéric Lopez, Jerzy Czaplicki, Pascal Demange, Alain MilonAbstract:THAP1, the founding member of a previously uncharacterized large family of cellular proteins (THAP proteins), is a sequence-specific DNA-binding factor that has recently been shown to regulate cell proliferation through modulation of pRb/E2F cell cycle target genes. THAP1 shares its DNA-binding THAP zinc finger domain with Drosophila P element transposase, zebrafish E2F6, and several nematode proteins interacting genetically with the retinoblastoma protein pRb. In this study, we report the three-dimensional structure and structure-function relationships of the THAP zinc finger of human THAP1. Deletion mutagenesis and multidimensional NMR spectroscopy revealed that the THAP domain of THAP1 is an atypical zinc finger of ∼80 residues, distinguished by the presence between the C2CH zinc coordinating residues of a short antiparallel β-sheet interspersed by a long loop-helix-loop insertion. Alanine scanning mutagenesis of this loop-helix-loop motif resulted in the identification of a number of critical residues for DNA recognition. NMR chemical shift perturbation analysis was used to further characterize the residues involved in DNA binding. The combination of the mutagenesis and NMR data allowed the mapping of the DNA binding interface of the THAP zinc finger to a highly positively charged area harboring multiple lysine and arginine residues. Together, these data represent the first structure-function analysis of a functional THAP domain, with demonstrated sequence-specific DNA binding activity. They also provide a structural framework for understanding DNA recognition by this atypical zinc finger, which defines a novel family of cellular factors linked to cell proliferation and pRb/E2F cell cycle pathways in humans, fish, and nematodes.
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structure function analysis of the thap zinc finger of THAP1 a large c2ch dna binding module linked to rb e2f pathways
Journal of Biological Chemistry, 2008Co-Authors: Damien Bessiere, Chrystelle Lacroix, Vincent Ecochard, Sébastien Campagne, Valérie Guillet, Lionel Mourey, Frédéric Lopez, Jerzy Czaplicki, Pascal Demange, Alain MilonAbstract:THAP1, the founding member of a previously uncharacterized large family of cellular proteins (THAP proteins), is a sequence-specific DNA-binding factor that has recently been shown to regulate cell proliferation through modulation of pRb/E2F cell cycle target genes. THAP1 shares its DNA-binding THAP zinc finger domain with Drosophila P element transposase, zebrafish E2F6, and several nematode proteins interacting genetically with the retinoblastoma protein pRb. In this study, we report the three-dimensional structure and structure-function relationships of the THAP zinc finger of human THAP1. Deletion mutagenesis and multidimensional NMR spectroscopy revealed that the THAP domain of THAP1 is an atypical zinc finger of approximately 80 residues, distinguished by the presence between the C2CH zinc coordinating residues of a short antiparallel beta-sheet interspersed by a long loop-helix-loop insertion. Alanine scanning mutagenesis of this loop-helix-loop motif resulted in the identification of a number of critical residues for DNA recognition. NMR chemical shift perturbation analysis was used to further characterize the residues involved in DNA binding. The combination of the mutagenesis and NMR data allowed the mapping of the DNA binding interface of the THAP zinc finger to a highly positively charged area harboring multiple lysine and arginine residues. Together, these data represent the first structure-function analysis of a functional THAP domain, with demonstrated sequence-specific DNA binding activity. They also provide a structural framework for understanding DNA recognition by this atypical zinc finger, which defines a novel family of cellular factors linked to cell proliferation and pRb/E2F cell cycle pathways in humans, fish, and nematodes.
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the thap zinc finger protein THAP1 regulates endothelial cell proliferation through modulation of prb e2f cell cycle target genes
Blood, 2007Co-Authors: Corinne Cayrol, Chrystelle Lacroix, Catherine Mathe, Vincent Ecochard, Vladimir Lazar, Emilie Loreau, Michele Ceribelli, Roberto Mantovani, Philippe Dessen, Luc AguilarAbstract:We recently cloned a novel human nuclear factor (designated THAP1) from postcapillary venule endothelial cells (ECs) that contains a DNA-binding THAP domain, shared with zebrafish E2F6 and several Caenorhabditis elegans proteins interacting genetically with retinoblastoma gene product (pRB). Here, we show that THAP1 is a physiologic regulator of EC proliferation and cell-cycle progression, 2 essential processes for angiogenesis. Retroviral-mediated gene transfer of THAP1 into primary human ECs inhibited proliferation, and large-scale expression profiling with microarrays revealed that THAP1-mediated growth inhibition is due to coordinated repression of pRB/E2F cell-cycle target genes. Silencing of endogenous THAP1 through RNA interference similarly inhibited EC proliferation and G1/S cell-cycle progression, and resulted in down-regulation of several pRB/E2F cell-cycle target genes, including RRM1, a gene required for S-phase DNA synthesis. Chromatin immunoprecipitation assays in proliferating ECs showed that endogenous THAP1 associates in vivo with a consensus THAP1-binding site found in the RRM1 promoter, indicating that RRM1 is a direct transcriptional target of THAP1. The similar phenotypes observed after THAP1 overexpression and silencing suggest that an optimal range of THAP1 expression is essential for EC proliferation. Together, these data provide the first links in mammals among THAP proteins, cell proliferation, and pRB/E2F cell-cycle pathways.
Tania Fuchs - One of the best experts on this subject based on the ideXlab platform.
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abnormalities of motor function transcription and cerebellar structure in mouse models of THAP1 dystonia
Human Molecular Genetics, 2015Co-Authors: Marta Ruiz, Andrika Morant, Jessica Kottwitz, Georgina Perezgarcia, Maitane Ortizvirumbrales, Aurelie Meneret, Tania Fuchs, Justine Bonet, Pedro Gonzalezalegre, Laurie J OzeliusAbstract:DYT6 dystonia is caused by mutations in THAP1 [Thanatos-associated (THAP) domain-containing apoptosis-associated protein] and is autosomal dominant and partially penetrant. Like other genetic primary dystonias, DYT6 patients have no characteristic neuropathology, and mechanisms by which mutations in THAP1 cause dystonia are unknown. THAP1 is a zinc-finger transcription factor, and most pathogenic THAP1 mutations are missense and are located in the DNA-binding domain. There are also nonsense mutations, which act as the equivalent of a null allele because they result in the generation of small mRNA species that are likely rapidly degraded via nonsense-mediated decay. The function of THAP1 in neurons is unknown, but there is a unique, neuronal 50-kDa THAP1 species, and THAP1 levels are auto-regulated on the mRNA level. Herein, we present the first characterization of two mouse models of DYT6, including a pathogenic knockin mutation, C54Y and a null mutation. Alterations in motor behaviors, transcription and brain structure are demonstrated. The projection neurons of the deep cerebellar nuclei are especially altered. Abnormalities vary according to genotype, sex, age and/or brain region, but importantly, overlap with those of other dystonia mouse models. These data highlight the similarities and differences in age- and cell-specific effects of a THAP1 mutation, indicating that the pathophysiology of THAP1 mutations should be assayed at multiple ages and neuronal types and support the notion of final common pathways in the pathophysiology of dystonia arising from disparate mutations.
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Screening of Brazilian families with primary dystonia reveals a novel THAP1 mutation and a de novo TOR1A GAG deletion
Movement disorders : official journal of the Movement Disorder Society, 2010Co-Authors: Patricia De Carvalho Aguiar, Tania Fuchs, Vanderci Borges, Kay Marie Lamar, Sonia M. A. Silva, Henrique Ballalai Ferraz, Laurie J OzeliusAbstract:The TOR1A and THAP1 genes were screened for mutations in a cohort of 21 Brazilian patients with Primary torsion dystonia (PTD). We identified a de novo delGAG mutation in the TOR1A gene in a patient with a typical DYT1 phenotype and a novel c.1A > G (p.Met1?) mutation in THAP1 in a patient with early onset generalized dystonia with speech involvement. Mutations in these two known PTD genes, TOR1A and THAP1, are responsible for about 10% of the PTD cases in our Brazilian cohort suggesting genetic heterogeneity and supporting the role of other genes in PTD. © 2010 Movement Disorder Society
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Direct interaction between causative genes of DYT1 and DYT6 primary dystonia.
Annals of neurology, 2010Co-Authors: Sophie Gavarini, Corinne Cayrol, Jean-philippe Girard, Michelle E Ehrlich, Tania Fuchs, Natalia Lyons, Laurie J OzeliusAbstract:Primary dystonia is a movement disorder characterized by sustained muscle contractions and in which dystonia is the only or predominant clinical feature. TOR1A(DYT1) and the transcription factor THAP1(DYT6) are the only genes identified thus far for primary dystonia. Using electromobility shift assays and chromatin immunoprecipitation (ChIP) quantitative polymerase chain reaction (qPCR), we demonstrate a physical interaction between THAP1 and the TOR1A promoter that is abolished by pathophysiologic mutations. Our findings provide the first evidence that causative genes for primary dystonia intersect in a common pathway and raise the possibility of developing novel therapies targeting this pathway.
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mutations in THAP1 dyt6 and generalised dystonia with prominent spasmodic dysphonia a genetic screening study
Lancet Neurology, 2009Co-Authors: Ana Djarmati, Katja Lohmann, Susanne A. Schneider, Tania Fuchs, Simone Zittel, Norbert Brüggemann, Susen Winkler, Heike Pawlack, Johann Hagenah, Aleksandar RakovicAbstract:Summary Background DYT6 is a primary, early-onset torsion dystonia; however, unlike in DYT1 dystonia, the symptoms of DYT6 dystonia frequently involve the craniocervical region. Recently, two mutations in THAP1 , the gene that encodes THAP (thanatos-associated protein) domain-containing apoptosis-associated protein 1 (THAP1), have been identified as a cause of DYT6 dystonia. Methods We screened THAP1 by sequence analysis and quantitative real-time polymerase chain reaction (PCR) in 160 white patients of European ancestry who had dystonia with an early age at onset (n=64), generalised dystonia (n=35), a positive family history of dystonia (n=56), or facial or laryngeal dystonia. Another 160 patients with dystonia were screened for reported and novel variants in THAP1 . 280 neurologically healthy controls were screened for the newly identified and previously reported changes in THAP1 and these and an additional 75 controls were screened for a rare non-coding mutation. Findings We identified two mutations in THAP1 (388_389delTC and 474delA), respectively, in two (1%) German patients from the 160 patients with dystonia. Both mutation carriers had laryngeal dystonia that started in childhood and both went on to develop generalised dystonia. Thus, two of three patients with early-onset generalised dystonia with orobulbar involvement had mutations in THAP1 . One of the identified patients with DYT6 dystonia had two family members with subtle motor signs who also carried the same mutation. A rare substitution in the 5′untranslated region (−236_235GA→TT) was found in 20 of 320 patients and in seven of 355 controls (p=0·0054). Interpretation Although mutations in THAP1 might have only a minor role in patients with different, but mainly focal, forms of dystonia, they do seem to be associated with early-onset generalised dystonia with spasmodic dysphonia. This combination of symptoms might be a characteristic feature of DYT6 dystonia and could be useful in the differential diagnosis of DYT1, DYT4, DYT12 , and DYT17 dystonia. In addition to the identified mutations, a rare non-coding substitution in THAP1 might increase the risk of dystonia. Funding Deutsche Forschungsgemeinschaft; Volkswagen Foundation; Dystonia Medical Research Foundation; University of Lubeck.
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mutations in THAP1 dyt6 in early onset dystonia a genetic screening study
Lancet Neurology, 2009Co-Authors: Laurie J Ozelius, Tania Fuchs, Deborah Raymond, Susan B Bressman, Gary A Heiman, Rachel SaunderspullmanAbstract:Summary Background Mutations in THAP1 were recently identified as the cause of DYT6 primary dystonia; a founder mutation was detected in Amish–Mennonite families, and a different mutation was identified in another family of European descent. To assess more broadly the role of this gene, we screened for mutations in families that included one family member who had early-onset, non-focal primary dystonia. Methods We identified 36 non- DYT1 multiplex families in which at least one person had non-focal involvement at an age of onset that was younger than 22 years. All three coding exons of THAP1 were sequenced, and the clinical features of individuals with mutations were compared with those of individuals who were negative for mutations in THAP1 . Genotype–phenotype differences were also assessed. Findings Of 36 families, nine (25%) had members with mutations in THAP1 , and most were of German, Irish, or Italian ancestry. One family had the Amish–Mennonite founder mutation, whereas the other eight families each had novel, potentially truncating or missense mutations. The clinical features of the families with mutations conformed to the previously described DYT6 phenotype; however, age at onset was extended from 38 years to 49 years. Compared with non-carriers, mutation carriers were younger at onset and their dystonia was more likely to begin in brachial, rather than cervical, muscles, become generalised, and include speech involvement. Genotype–phenotype differences were not found. Interpretation Mutations in THAP1 underlie a substantial proportion of early-onset primary dystonia in non- DYT1 families. The clinical features that are characteristic of affected individuals who have mutations in THAP1 include limb and cranial muscle involvement, and speech is often affected. Funding Dystonia Medical Research Foundation; Bachmann–Strauss Dystonia and Parkinson Foundation; National Institute of Neurological Disorders and Stroke; Aaron Aronov Family Foundation.