RESUMO
Spinocerebellar ataxia type 17 or ATX-TBP is a CAG/CAA repeat expansion disorder characterized by marked clinical heterogeneity. Reports of affected carriers with subthreshold repeat expansions and of patients with Parkinson's disease (PD) with expanded repeats have cast doubt on the established cutoff values of the expansions and the phenotypic spectrum of this disorder. The objective of this systematic review was to explore the genotype-phenotype relationships for repeat expansions in TBP to delineate the ATX-TBP phenotype and reevaluate the pathological range of repeat expansions. The International Parkinson and Movement Disorder Society Genetic Mutation Database (MDSGene) standardized data extraction protocol was followed. Clinically affected carriers of reported ATX-TBP expansions were included. Publications that contained repeat sizes in screened cohorts of patients with PD and/or healthy individuals were included for a separate evaluation of cutoff values. Phenotypic and genotypic data for 346 ATX-TBP patients were curated. Overall, 97.7% of the patients had ≥41 repeats, while 99.6% of patients with PD and 99.9% of healthy individuals had ≤42 repeats, with a gray zone of reduced penetrance between 41 and 45 repeats. Pure parkinsonism was more common in ATX-TBP patients with 41 to 45 repeats than in the group with ≥46 repeats, which conversely more often presented with a complex phenotype with mixed movement disorders. An updated genotype-phenotype assessment for ATX-TBP is provided, and new repeat expansion cutoff values of reduced penetrance (41-45 expanded repeats) and full penetrance (46-66 expanded repeats) are proposed. These adjusted cutoff values will have diagnostic and counseling implications and may guide future clinical trial protocol. © 2022 The Authors. Movement Disorders published by Wiley Periodicals LLC on behalf of International Parkinson and Movement Disorder Society.
Assuntos
Doença de Parkinson , Ataxias Espinocerebelares , Humanos , Estudos de Associação Genética , Doença de Parkinson/genética , Ataxias Espinocerebelares/genética , Proteína de Ligação a TATA-Box/genética , Expansão das Repetições de TrinucleotídeosRESUMO
The assembly of transcription complexes on eukaryotic promoters involves a series of steps, including chromatin remodeling, recruitment of TATA-binding protein (TBP)-containing complexes, the RNA polymerase II holoenzyme, and additional basal transcription factors. This review describes the transcriptional regulation by TBP and its corresponding homologs that constitute the TBP family and their interactions with promoter DNA. The C-terminal core domain of TBP is highly conserved and contains two structural repeats that fold into a saddle-like structure, essential for the interaction with the TATA-box on DNA. Based on the TBP C-terminal core domain similarity, three TBP-related factors (TRFs) or TBP-like factors (TBPLs) have been discovered in metazoans, TRF1, TBPL1, and TBPL2. TBP is autoregulated, and once bound to DNA, repressors such as Mot1 induce TBP to dissociate, while other factors such as NC2 and the NOT complex convert the active TBP/DNA complex into inactive, negatively regulating TBP. TFIIA antagonizes the TBP repressors but may be effective only in conjunction with the RNA polymerase II holoenzyme recruitment to the promoter by promoter-bound activators. TRF1 has been discovered inDrosophila melanogasterandAnophelesbut found absent in vertebrates and yeast. TBPL1 cannot bind to the TATA-box; instead, TBPL1 prefers binding to TATA-less promoters. However, TBPL1 shows a stronger association with TFIIA than TBP. The TCT core promoter element is present in most ribosomal protein genes inDrosophilaand humans, and TBPL1 is required for the transcription of these genes. TBP directly participates in the DNA repair mechanism, and TBPL1 mediates cell cycle arrest and apoptosis. TBPL2 is closely related to its TBP paralog, showing 95% sequence similarity with the TBP core domain. Like TBP, TBPL2 also binds to the TATA-box and shows interactions with TFIIA, TFIIB, and other basal transcription factors. Despite these advances, much remains to be explored in this family of transcription factors.
Assuntos
RNA Polimerase II , Proteína de Ligação a TATA-Box , Fatores de Transcrição , Transcrição Gênica , Adenosina Trifosfatases/genética , Animais , DNA/genética , Drosophila , Holoenzimas/genética , Holoenzimas/metabolismo , Humanos , Proteínas Nucleares/genética , RNA Polimerase II/genética , RNA Polimerase II/metabolismo , Saccharomyces cerevisiae/genética , Proteínas de Saccharomyces cerevisiae , TATA Box/genética , Proteínas Semelhantes à Proteína de Ligação a TATA-Box/química , Proteínas Semelhantes à Proteína de Ligação a TATA-Box/genética , Proteínas Semelhantes à Proteína de Ligação a TATA-Box/metabolismo , Fatores Associados à Proteína de Ligação a TATA , Proteína de Ligação a TATA-Box/química , Proteína de Ligação a TATA-Box/genética , Proteína de Ligação a TATA-Box/metabolismo , Fator de Transcrição TFIIA/genética , Fator de Transcrição TFIIA/metabolismo , Fatores de Transcrição/genéticaRESUMO
Huntington's disease (HD) is a genetic neurodegenerative progressive and fatal disease characterized by motor disorder, cognitive impairment, and behavioral problems, caused by expanded repeats of CAG trinucleotides in the HTT gene. The aim of this study was to investigate the influence of TBP gene CAG/CAA repeats in conjunction with HTT gene CAG repeats, on the age at HD onset in Brazilian individuals. Individuals diagnosed as molecularly negative for HD presented 29-39 TBP CAG/CAA. Their most frequent allele had 36 repeats. In individuals diagnosed as molecularly positive for HD, a range of 25-40 TBP CAG/ CAA was found. The most frequent TBP allele had 38 repeats. We also conducted TBP direct Sanger sequencing of some samples which demonstrated other four TBP structures different from the basic TBP structure and others reported in the literature. The HTT expanded CAG and TBP CAG/CAA repeat sizes jointly explained 66% of the age at onset (AO) in our HD patients. The strongest variable in the model associated with AO was the number of expanded HTT CAG repeats. The difference between the association of HD AO with HTT expanded CAG together with TBP CAG/CAA and the association of HD AO with HTT expanded CAG was 0.001 (∆R2). Therefore, we found a weak association (0.1%) of TBP CAG/CAA repeats on HD AO, if any.
Assuntos
Doença de Huntington , Idade de Início , Brasil , Humanos , Proteína Huntingtina/genética , Doença de Huntington/genética , Expansão das Repetições de TrinucleotídeosRESUMO
Transcription is the first step of gene expression regulation and is a fundamental mechanism for establishing the viability and development of a cell. The TATA box-binding protein (TBP) interaction with a TATA box in a promoter is one of the best studied mechanisms in transcription initiation. TBP is a transcription factor that is highly conserved from archaea to humans and is essential for the transcription initiated by each of the three RNA polymerases. In addition, the discovery of TBP-related factor 1 (TRF1) and other factors related to TBP shed light on the variability among transcription initiation complexes, thus demonstrating that the compositions of these complexes are, in fact, more complicated than originally believed. Despite these facts, the majority of studies on transcription have been performed on animal, plant and fungal cells, which serve as canonical models, and information regarding protist cells is relatively scarce. The aim of this work is to review the diversity of the TBPs that have been documented in protists and describe some of the specific features that differentiate them from their counterparts in higher eukaryotes.
Assuntos
Eucariotos/genética , TATA Box , Proteína de Ligação a TATA-Box , Transcrição Gênica , Eucariotos/metabolismo , Genes de Protozoários , Variação Genética , Giardia/genética , Giardia/metabolismo , Leishmania/genética , Leishmania/metabolismo , Filogenia , Proteínas de Protozoários/genética , Proteínas de Protozoários/metabolismo , Proteína de Ligação a TATA-Box/genética , Proteína de Ligação a TATA-Box/metabolismo , Trypanosoma/genética , Trypanosoma/metabolismoRESUMO
The protozoan parasite Trichomonas vaginalis is a common human pathogen from one of the earliest-diverging eukaryotic lineages. At the transcriptional level, the highly conserved Inr element of RNA pol II-transcribed genes surrounds the transcription start site and is recognised by IBP39, a protein exclusive of T. vaginalis. Typical TATA boxes have not been identified in this organism but, in contrast, BLAST analyses of the T. vaginalis genome identified two genes encoding putative TATA-binding proteins (herein referred to as TvTBP1 and TvTBP2). The goal of this work was to characterise these two proteins at the molecular level. Our results show that both TvTBPs theoretically adopt the saddle-shaped structure distinctive to TBPs and both Tvtbp genes are expressed in T. vaginalis. TvTBP1 did not complement a Saccharomyces cerevisiae mutant lacking TBP; however, TvTBP1 and TvTBP2 proteins bound T. vaginalis DNA promoter sequences in EMSA assays. We propose that TvTBP1 may be part of the preinitiation transcription complex in T. vaginalis since TvTBP1 recombinant protein was able to bind IBP39 in vitro. This work represents the first approach towards the characterisation of general transcription factors in this early divergent organism.
Assuntos
Proteínas de Ligação a DNA/metabolismo , Proteínas de Protozoários/metabolismo , Proteína de Ligação a TATA-Box/metabolismo , Trichomonas vaginalis/metabolismo , Modelos Moleculares , Regiões Promotoras Genéticas , Ligação Proteica , Proteínas de Protozoários/química , Proteínas de Protozoários/genética , Proteínas Recombinantes/química , Proteínas Recombinantes/genética , Proteínas Recombinantes/metabolismo , Saccharomyces cerevisiae/genética , Proteína de Ligação a TATA-Box/química , Proteína de Ligação a TATA-Box/genética , Transcrição Gênica , Trichomonas vaginalis/genéticaRESUMO
El desarrollo de generadores radisotópicos solo para uso industrial y de radiotrazadores, a partir de los ya existentes como el de 
/
, se ha potenciado en los últimos 10 años como una opción atractiva ante las dificultades de garantizar la disponibilidad de radiotrazadores para aplicaciones en la industria. Teniendo en cuenta que la extracción con la mezcla 30 % TBF-16 % TOA/ciclohexano se utilizó con éxito para adecuar el eluido del generador de / como radiotrazador de fluidos orgánicos, se realizó un estudio de optimización de la composición volumétrica de esta mezcla; se estableció un modelo matemático para predecir el grado de extracción (R %) del , en dependencia de las concentraciones volumétricas de TOA y TBF, para una actividad de 3.1 MBq, y se determinó que, aún reduciendo la concentración volumétrica de TBF al 1 % y la de TOA al 0.3 %, se extrajo el 96.44 ± 0.21 % del 
.
The development of radioisotope generators for industrial use, and of radiotracers from those already existing as that of /, has been strengthened in the last 10 years like an attractive option to solve the difficulties in guaranteeing the availability of radiotracers for application in the industry. Extraction with the mixture 30 % TBP-16 % TOA/cyclohexane has been successfully used to adapt the , eluted from / generator, as an organic fluid radiotracer. This work presents an optimization study of volumetric composition of this mixture, to guide the extraction process towards the best cost benefit relation according to necessary activity at a given application. Based on experimental results, a model that predicts extraction yield (R %) as a dependent variable of TOA and TBP volumetric concentrations was established, for 3.1 MBq of . Moreover, the outcomes show that even when TBP volumetric concentration decreases from 30 to 1 % and TOA concentration, from 16 to 0.3 %, the extraction yield was 96.44 ± 0.21%.
RESUMO
An alternative for radiotracer technique in Cuba, is the use of sodium pertechnate (Na99mTcO4) eluted from 99Mo/99mTc generator. This radioisotope generator is produced by the Cuban Center of Isotopes (CENTIS). Taking into account that solvent extraction is an usual procedure to separate 99mTcO4- from the fission products, this technique was used to get a radiotracer in organic phase from the eluted 99mTcO4-, using 30% TBP-16% TOA/ n-dodecane and 30% TBP-16% TOA/ ciclohexane as solvents. On the other hand, MEK is used as solvent in the so called 99Mo/99mTc extraction radioisotope generator. Consequently, it was also evaluated. In this paper, the influence on the extraction degree, of the volumetric ratio between organic and aqueous phases, the time of phase shaking and the time elapsed between shaking-end and phase separation were studied. Furthermore, in the case of MEK, the influence of NaOH concentration in the aqueous phase was considered. For the extraction using 30% TBP-16% TOA/ciclohexane and 30% TBP-16% TOA/n-dodecane, the influence of pH was also studied. The extraction degree of 99TcO4- eluted from generator using MEK, as well as 30% TBP-16% TOA in n-dodecane or ciclohexane, in adequate conditions, was higher than 99%, so, any of the tested solvents could be used to get a 99mTc radiotracer in organic phase.
Una alternativa para el empleo de la técnica de radiotrazadores en Cuba ha sido el uso del pertecnetato de sodio (Na99TcO4) eluido del generador de 99Mo/99Tc producido en el Centro de Isótopos. Teniendo en cuenta que la extracción se utiliza regularmente para separar al 99mTcO4- de los productos de fisión, se evaluó este método para obtener un radiotrazador en fase orgánica a partir del 99TcO4-, empleando como solventes la mezcla 30% TBF-16% TOA en n-dodecano y en ciclohexano. Por otro lado, la metiletilcetona se utiliza como solvente en el generador radisotópico de extracción de 99Mo/99mTc, por lo que también se evaluó para estos fines. En el trabajo se estudió la influencia de la relación volumétrica de la fase acuosa y la orgánica, del tiempo de contacto en agitación y del tiempo de reposo antes de la separación de las fases, en el grado de extracción del 99mTc. En el caso de la metiletilcetona se evaluó además, la influencia de la concentración de NaOH en la fase acuosa y para la extracción, con 30% TBF-16% TOA en ciclohexano y en n-dodecano, la del pH. El grado de extracción del 99TcO4- eluido del generador empleando, ya sea la metiletlicetona o la mezcla 30% TBF-16% TOA en n-dodecano o ciclohexano, en condiciones adecuadas, fue superior al 99 %; por tanto, cualquiera de los solventes evaluados se puede utilizar para obtener radiotrazadores de 99mTc en fase orgánica.