Control of Ubp3 ubiquitin protease activity by the Hog1 SAPK modulates transcription upon osmostress

Here, the Hog1 kinase interacts with and activates the ubiquitin protease Ubp3 in a stress-dependent manner. The phosphorylation of Ubp3 enhances RNA polymerase II occupancy on osmotic stress-responsive genes

Ask yeast how to burn your fats: lessons learned from the metabolic adaptation to salt stress

[EN] Here, we review and update the recent advances in the metabolic control during the adaptive response of budding yeast to hyperosmotic and salt stress, which is one of the best understood signaling events at the molecular level. This environmental stress can be easily applied and hence has been exploited in the past to generate an impressively detailed and comprehensive model of cellular adaptation. It is clear now that this stress modulates a great number of different physiological functions of the cell, which altogether contribute to cellular survival and adaptation. Primary defense mechanisms are the massive induction of stress tolerance genes in the nucleus, the activation of cation transport at the plasma membrane, or the production and intracellular accumulation of osmolytes. At the same time and in a coordinated manner, the cell shuts down the expression of housekeeping genes, delays the progression of the cell cycle, inhibits genomic replication, and modulates translation efficiency to optimize the response and to avoid cellular damage. To this fascinating interplay of cellular functions directly regulated by the stress, we have to add yet another layer of control, which is physiologically relevant for stress tolerance. Salt stress induces an immediate metabolic readjustment, which includes the up-regulation of peroxisomal biomass and activity in a coordinated manner with the reinforcement of mitochondrial respiratory metabolism. Our recent findings are consistent with a model, where salt stress triggers a metabolic shift from fermentation to respiration fueled by the enhanced peroxisomal oxidation of fatty acids. 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Coordinated gene regulation in the initial phase of salt stress adaptation

This research was originally published in Journal of Biological Chemistry, 2015 - 16 : 10175- 10163 © the American Society for Biochemistry and Molecular Biology[EN] Stress triggers complex transcriptional responses, which include both gene activation and repression. We used time-resolved reporter assays in living yeast cells to gain insights into the coordination of positive and negative control of gene expression upon salt stress. We found that the repression of housekeeping genes coincides with the transient activation of defense genes and that the timing of this expression pattern depends on the severity of the stress. Moreover, we identified mutants that caused an alteration in the kinetics of this transcriptional control. Loss of function of the vacuolar H+-ATPase (vma1) or a defect in the biosynthesis of the osmolyte glycerol (gpd1) caused a prolonged repression of housekeeping genes and a delay in gene activation at inducible loci. Both mutants have a defect in the relocation of RNA polymerase II complexes at stress defense genes. Accordingly salt-activated transcription is delayed and less efficient upon partially respiratory growth conditions in which glycerol production is significantly reduced. Furthermore, the loss of Hog1 MAP kinase function aggravates the loss of RNA polymerase II from housekeeping loci, which apparently do not accumulate at inducible genes. Additionally the Def1 RNA polymerase II degradation factor, but not a high pool of nuclear polymerase II complexes, is needed for efficient stress-induced gene activation. The data presented here indicate that the finely tuned transcriptional control upon salt stress is dependent on physiological functions of the cell, such as the intracellular ion balance, the protective accumulation of osmolyte molecules, and the RNA polymerase II turnover.This work was supported by Ministerio de Economia y Competitividad Grant BFU2011-23326 (to M. 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Aplicación de nuevos procesos multicomponente a la síntesis de derivados de quinolina

Las principales transformaciones sintéticas desarrolladas en este trabajo fueron: 1. Un nuevo proceso multicomponente para la síntesis de trans‐4‐anilino‐2‐ aril‐1,2,3,4-‐tetrahidroquinolinas y estudio de su aromatización. 2. Transformación de 4‐alquil‐4-‐viniltetrahidroquinolinas en quinolinas, por transposición C4‐C3 o pérdida de una cadena olefínica 3. Uso de 2‐acil‐4‐hidrazono‐1,2,3,4‐tetrahidroquinolinas para la obtención diastereoselectiva de derivados de 2,6‐metanobenzo[e][1,4]diazocina. 4. Desarrollo de la reacción de nitro-Mannich intramolecular para la síntesis diastereoselectiva de tetrahidroquinolinas sustituidas en 2 y 3. 5. Desarrollo del nitrato cérico amónico como catalizador para la reacción de Friedländer‐Borsche y su aplicación a la síntesis de luotoninas. [ABSTRACT]The main synthetic transformations developed in this work were: 1. A new multicomponent process for the synthesis of trans‐4‐anilino‐2‐aryl‐ 1,2,3,4-‐tetrahydroquinolines and studies on their aromatization. 2. Transformation of 4-alkyl­4‐vinyltetrahydroquinolines into quinolines, by C4 to C3 shift or loss of their vinyl side chain. 3. Use of 2‐acyl‐4‐hydrazono‐1,2,3,4‐tetrahydroquinolines for the diastereoselective preparation of 2,6‐metanobenzo[e][1,4]diazocines. 4. Development of the intramolecular nitro-Mannich reaction as a diastereoselective route to 2,3-‐disubstitutes tetrahydroquinolines. 5. Development of ceric ammonium nitrate as a catalyst for the Friedländer‐ Borsche reaction and its application for luotonin synthesis

Synthesis of Polysubstituted, Functionalized Quinolines through a Metal-Free Domino Process Involving a C₄–C₃ Functional Group Rearrangement

4-Alkyl-1,2,3,4-tetrahydroquinolines bearing a 4-vinyl unit ending in an electron-withdrawing group were efficiently transformed into polysubstituted, C₃-functionalized quinolines upon heating in refluxing <i>o</i>-dichlorobenzene, in a domino reaction involving an unusual C₄–C₃ functional group rearrangement

Different Pathological Complete Response Rates According to PAM50 Subtype in HER2+ Breast Cancer Patients Treated With Neoadjuvant Pertuzumab/Trastuzumab vs. Trastuzumab Plus Standard Chemotherapy: An Analysis of Real-World Data.

Background: Double blockade with pertuzumab and trastuzumab combined with chemotherapy is the standard neoadjuvant treatment for HER2-positive early breast cancer. Data derived from clinical trials indicates that the response rates differ among intrinsic subtypes of breast cancer. The aim of this study is to determine if these results are valid in real-world patients. Methods: A total of 259 patients treated in eight Spanish hospitals were included and divided into two cohorts: Cohort A (132 patients) received trastuzumab plus standard neoadjuvant chemotherapy (NAC), and Cohort B received pertuzumab and trastuzumab plus NAC (122 patients). Pathological complete response (pCR) was defined as the complete disappearance of invasive tumor cells. Assignment of the intrinsic subtype was realized using the research-based PAM50 signature. Results: There were more HER2-enriched tumors in Cohort A (70 vs. 56%) and more basal-like tumors in Cohort B (12 vs. 2%), with similar luminal cases in both cohorts (luminal A 12 vs. 14%; luminal B 14 vs. 18%). The overall pCR rate was 39% in Cohort A and 61% in Cohort B. Better pCR rates with pertuzumab plus trastuzumab than with trastuzumab alone were also observed in all intrinsic subtypes (luminal PAM50 41 vs. 11.4% and HER2-enriched subtype 73.5 vs. 50%) but not in basal-like tumors (53.3 vs. 50%). In multivariate analysis the only significant variables related to pCR in both luminal PAM50 and HER2-enriched subtypes were treatment with pertuzumab plus trastuzumab (Cohort B) and histological grade 3. Conclusions: With data obtained from patients treated in clinical practice, it has been possible to verify that the addition of pertuzumab to trastuzumab and neoadjuvant chemotherapy substantially increases the rate of pCR, especially in the HER2-enriched subtype but also in luminal subtypes, with no apparent benefit in basal-like tumors

B-Ring-Aryl Substituted Luotonin A Analogues with a New Binding Mode to the Topoisomerase 1-DNA Complex Show Enhanced Cytotoxic Activity

<div><p>Topoisomerase 1 inhibition is an important strategy in targeted cancer chemotherapy. The drugs currently in use acting on this enzyme belong to the family of the camptothecins, and suffer severe limitations because of their low stability, which is associated with the hydrolysis of the δ-lactone moiety in their E ring. Luotonin A is a natural camptothecin analogue that lacks this functional group and therefore shows a much-improved stability, but at the cost of a lower activity. Therefore, the development of luotonin A analogues with an increased potency is important for progress in this area. In the present paper, a small library of luotonin A analogues modified at their A and B rings was generated by cerium(IV) ammonium nitrate-catalyzed Friedländer reactions. All analogues showed an activity similar or higher than the natural luotonin A in terms of topoisomerase 1 inhibition and some compounds had an activity comparable to that of camptothecin. Furthermore, most compounds showed a better activity than luotonin A in cell cytotoxicity assays. In order to rationalize these results, the first docking studies of luotonin-topoisomerase 1-DNA ternary complexes were undertaken. Most compounds bound in a manner similar to luotonin A and to standard topoisomerase poisons such as topotecan but, interestingly, the two most promising analogues, bearing a 3,5-dimethylphenyl substituent at ring B, docked in a different orientation. This binding mode allows the hydrophobic moiety to be shielded from the aqueous environment by being buried between the deoxyribose belonging to the G(+1) guanine and Arg364 in the scissile strand and the surface of the protein and a hydrogen bond between the D-ring carbonyl and the basic amino acid. The discovery of this new binding mode and its associated higher inhibitory potency is a significant advance in the design of new topoisomerase 1 inhibitors.</p></div

B-ring substitution improves hydrogen bonding to Arg364.

<p><b>A.</b> Overlay of luotonin A (<b>3a</b>) and compounds <b>3b</b>–<b>e</b> at their binding sites, showing that the presence of the substituent at the C-14 position of ring B induces a rotation of the docking pose that leads to improved hydrogen bonding to Arg364 and hence to a better fit to the binding site. <b>B.</b> Docking of luotonin A showing the position of Asn352, which would interfere with any ring-B substituent. <b>C.</b> Docking of the B-phenyl substituted compound <b>3c,</b> also showing the position of Asn352.</p