Procesamiento proteolítico y regulación de la localización subcelular del factor de transcripción PacC por el PH ambiental

Tesis de la Universidad Complutense de Madrid, Facultad de Farmacia, Departamento de Bioquímica y Biología Molecular II, leída el 02-11-1999En el trabajo realizado en esta Tesis Doctoral se han estudiado diferentes aspectos de factores de transcripción PacC(factor que media la regulación por pH de la espresión génica en Aspergillus nidulans) en respuesta a un pH ambiental alcalino. Se ha demostrado que el producto primario de traducción de pacC se procesa proteolíticamente para dar lugar a una forma activa de la proteína mucho más estable y que la ruta de transducción de la señal de pH ambiental alcalino codificada por los genes pal, es necesaria para que este procesamiento pueda tener lugar; se ha caracterizado el principal punto de corte de la proteasa procesativa y se han obtenido evidencias muy consistentes que favorecen la hipótesis de que el pH ambiental, más que regular la actividad de la proteasa, regula la accesibilidad de PacC a esta proteasa. Así mismo se ha comprobado que el pH ambiental regula la localización nuclear de PacC; que, a diferencia de lo que ocurre con el producto primario de traducción, la localización nuclear de la forma procesada de PacC es independiente de la funcionalidad de la ruta pa1 y que la región de PacC que contiene los dedos de zinc es necesaria para que proteínas PacC similares en tamaño a la forma procesada de la proteína se localicen en el núcleo. Finalmente lo resultados obtenidos en esta tesis muestran que pacC no regula positivamente su propia expresión y que el producto primario de traducción de pacC no es inactivo y parece modular negativamente la función pacCSección Deptal. de Bioquímica y Biología Molecular (Farmacia)Fac. de FarmaciaTRUEpu

eEF1A mediates the nuclear export of SNAG-containing proteins via the exportin5-aminoacyl-tRNA complex

This is an open-access article distributed under the terms of the Creative Commons Attribution-NonCommercial-No Derivative Works License.Exportin5 mediates the nuclear export of double-stranded RNAs, including pre-microRNAs, adenoviral RNAs, and tRNAs. When tRNAs are aminoacylated, the Exportin5-aminoacyl (aa)-tRNA complex recruits and coexports the translation elongation factor eEF1A. Here, we show that eEF1A binds to Snail transcription factors when bound to their main target, the E-cadherin promoter, facilitating their export to the cytoplasm in association with the aa-tRNA-Exportin5 complex. Snail binds to eEF1A through the SNAG domain, a protein nuclear export signal present in several transcription factor families, and this binding is regulated by phosphorylation. Thus, we describe a nuclear role for eEF1A and provide a mechanism for protein nuclear export that attenuates the activity of SNAG-containing transcription factors. © 2013 The Authors.This work was supported by grants from the Spanish Ministry of Science and Innovation (SAF2010-21143 to A.C.; BFU2008-01042 and CONSOLIDER-INGENIO 2010 CSD2007-00023 to M.A.N.; and CDS2007-00017 to A.C. and M.A.N.’s lab) and the Generalitat Valenciana (Prometeo 2008/049 and PROMETEOII/2013/002 to M.A.N.).Peer Reviewe

eEF1A Mediates the Nuclear Export of SNAG-Containing Proteins via the Exportin5-Aminoacyl-tRNA Complex

Erratum: (Cell Reports 5, 727–737; November 14, 2013) In this article, the two lanes in Figure 4E were inadvertently used twice when making the composite. The correct version of Figure 4 appears her

Real-world assessment and characteristics of men with benign prostatic hyperplasia (BPH) in primary care and urology clinics in Spain

Objectives: To describe the real-world demographic and clinical characteristics of patients with lower urinary tract symptoms (LUTS) as a result of benign prostatic hyperplasia (BPH) in Spain. Methodology: This observational, retrospective, multicentre study conducted in primary care and urology clinics in Spain included men aged ≥50 years diagnosed (≤8 years prior to study visit) with LUTS caused by BPH. The primary endpoint was demographic and clinical characteristics; secondary endpoints included disease progression and diagnostic tests across both healthcare settings. Results: A total of 670 patients were included (primary care: n = 435; urology: n = 235). Most patients had moderate/severe LUTS (74.6%) and prostate volume >30 cc (81.7%), with no differences between settings. More patients had prostate-specific antigen (PSA) ≥1.5 ng/mL in primary care (74.5%) versus urology (67.7%). Progression criteria were prevalent (48.9%). Clinical criteria were more commonly used than the International Prostate Symptom Score (IPSS) to evaluate LUTS at diagnosis (primary care: clinical criteria 73.0%; IPSS: 26.9%; urology: clinical criteria 76.5%; IPSS: 23.4%). Proportion of patients with moderate/severe LUTS at diagnosis was lower using clinical criteria than IPSS, and the proportion of patients with 'worsening' LUTS (diagnosis to study visit) was higher when using clinical criteria versus IPSS. In both healthcare settings, the most commonly used diagnostic tests were general and urological clinical history and PSA. Conclusion: Demographic and clinical characteristics of patients with BPH in Spain were similar in primary care and urology; however, assessment criteria to evaluate LUTS severity differ and are not completely aligned with clinical guideline recommendations. Increased use of recommended assessments may enhance optimal BPH management

Procesamiento proteolítico y regulación de la localización subcelular del factor de transcripción PacC por el PH ambiental

En el trabajo realizado en esta Tesis Doctoral se han estudiado diferentes aspectos de factores de transcripción PacC(factor que media la regulación por pH de la espresión génica en Aspergillus nidulans) en respuesta a un pH ambiental alcalino. Se ha demostrado que el producto primario de traducción de pacC se procesa proteolíticamente para dar lugar a una forma activa de la proteína mucho más estable y que la ruta de transducción de la señal de pH ambiental alcalino codificada por los genes pal, es necesaria para que este procesamiento pueda tener lugar; se ha caracterizado el principal punto de corte de la proteasa procesativa y se han obtenido evidencias muy consistentes que favorecen la hipótesis de que el pH ambiental, más que regular la actividad de la proteasa, regula la accesibilidad de PacC a esta proteasa. Así mismo se ha comprobado que el pH ambiental regula la localización nuclear de PacC; que, a diferencia de lo que ocurre con el producto primario de traducción, la localización nuclear de la forma procesada de PacC es independiente de la funcionalidad de la ruta pa1 y que la región de PacC que contiene los dedos de zinc es necesaria para que proteínas PacC similares en tamaño a la forma procesada de la proteína se localicen en el núcleo. Finalmente lo resultados obtenidos en esta tesis muestran que pacC no regula positivamente su propia expresión y que el producto primario de traducción de pacC no es inactivo y parece modular negativamente la función pac

Reply: “Comment on Search and Selection of Probiotics that Improve Mucositis Symptoms in Oncologic Patients. A Systematic Review. Nutrients 2019, 11, 2322”

In the article, “Search and Selection of Probiotics That Improve Mucositis Symptoms in Oncologic Patients [...

Disruption of phacA, an Aspergillus nidulans gene encoding a novel cytochrome P450 monooxygenase catalyzing phenylacetate 2-hydroxylation, results in penicillin overproduction

7 p.-6 fig.Aspergillus nidulans utilizes phenylacetate as a carbon source via homogentisate, which is degraded to fumarate and acetoacetate. Mutational evidence strongly suggested that phenylacetate is converted to homogentisate through two sequential hydroxylating reactions in positions 2 and 5 of the aromatic ring. Using cDNA substraction techniques, we have characterized a gene, denoted phacA, whose transcription is strongly induced by phenylacetate and which putatively encodes a cytochrome P450 protein. A disrupted phacA strain does not grow on phenylacetate but grows on 2-hydroxy- or 2, 5-dihydroxyphenylacetate. Microsomal extracts of the disrupted strain are deficient in the NADPH-dependent conversion of phenylacetate to 2-hydroxyphenylacetate. We conclude that PhacA catalyzes the ortho-hydroxylation of phenylacetate, the first step of A. nidulans phenylacetate catabolism. The involvement of a P450 enzyme in the ortho-hydroxylation of a monoaromatic compound has no precedent. In addition, PhacA shows substantial sequence divergence with known cytochromes P450 and defines a new family of these enzymes, suggesting that saprophytic fungi may represent a source of novel cytochromes P450. Phenylacetate is a precursor for benzylpenicillin production. phacA disruption increases penicillin production 3-5-fold, indicating that catabolism competes with antibiotic biosynthesis for phenylacetate and strongly suggesting strategies for Penicillium chrysogenum strain improvement by reverse genetics.This work was supported by Spanish Comisión Interministerial de Ciencia y Tecnologı́a Grants BIO94-932 and BIO97-348 and Antibióticos S. A.Peer reviewe

Characterization of Snail nuclear import pathways as representatives of C2H2 zinc finger transcription factors

9 pages.-- PMID: 19386897 [PubMed].-- Printed version published May 1, 2009.Supporting information (Suppl. fig S1) available at: http://jcs.biologists.org/cgi/content/full/122/9/1452/DC1Snail proteins are C2H2 class zinc finger transcription factors involved in different processes during embryonic development, as well as in several adult pathologies including cancer and organ fibrosis. The expression of Snail transcription factors is tightly regulated at the transcriptional level and their activity is modulated by their subcellular localization. Given the importance of this gene family in physiology and pathology, it is essential to understand the mechanisms by which Snail proteins are imported into or exported out of the nucleus. Here we show that several importins mediate the nuclear import of the human Snail proteins and we identify a unique nuclear localization signal (NLS), recognized by all the importins, that has been conserved during the evolution of the Snail family. This NLS is characterized by the presence of basic residues at defined positions in at least three consecutive zinc fingers. Interestingly, the consensus residues for importin-binding are also involved in DNA binding, suggesting that importins could prevent non-specific binding of these transcription factors to cytoplasmic polyanions. Importantly, the identified basic residues are also conserved in other families of C2H2 transcription factors whose nuclear localization requires the zinc finger region.This work was supported by the Spanish Ministry of Education and Science (Grants BFU2005-05772, NAN2004-09230-C04-04 and CONSOLIDER-INGENIO 2010 CSD2007-00017) to M.A.N.; BIO2007-67304-C02-02 to J.M.S. and B.M. and a grant (GV06/027) to J.M.M., who holds a Ramon y Cajal Contract (Spanish Ministry of Science and Innovation).Peer reviewe