RESUMO
Heat acclimation improves the tolerance of organisms to severe heat stress. Our previous work showed that in Arabidopsis (Arabidopsis thaliana), the "memory" of heat acclimation treatment decayed faster in the absence of the heat-stress-associated 32-kD protein HSA32, a heat-induced protein predominantly found in plants. The HSA32 null mutant attains normal short-term acquired thermotolerance but is defective in long-term acquired thermotolerance. To further explore this phenomenon, we isolated Arabidopsis defective in long-term acquired thermotolerance (dlt) mutants using a forward genetic screen. Two recessive missense alleles, dlt1-1 and dlt1-2, encode the molecular chaperone heat shock protein101 (HSP101). Results of immunoblot analyses suggest that HSP101 enhances the translation of HSA32 during recovery after heat treatment, and in turn, HSA32 retards the decay of HSP101. The dlt1-1 mutation has little effect on HSP101 chaperone activity and thermotolerance function but compromises the regulation of HSA32. In contrast, dlt1-2 impairs the chaperone activity and thermotolerance function of HSP101 but not the regulation of HSA32. These results suggest that HSP101 has a dual function, which could be decoupled by the mutations. Pulse-chase analysis showed that HSP101 degraded faster in the absence of HSA32. The autophagic proteolysis inhibitor E-64d, but not the proteasome inhibitor MG132, inhibited the degradation of HSP101. Ectopic expression of HSA32 confirmed its effect on the decay of HSP101 at the posttranscriptional level and showed that HSA32 was not sufficient to confer long-term acquired thermotolerance when the HSP101 level was low. Taken together, we propose that a positive feedback loop between HSP101 and HSA32 at the protein level is a novel mechanism for prolonging the memory of heat acclimation.
Assuntos
Aclimatação/genética , Proteínas de Arabidopsis/metabolismo , Arabidopsis/genética , Arabidopsis/fisiologia , Proteínas de Choque Térmico/metabolismo , Temperatura Alta , Proteínas de Plantas/metabolismo , Fatores de Transcrição/metabolismo , Transcrição Gênica , Aclimatação/efeitos dos fármacos , Arabidopsis/efeitos dos fármacos , Autofagia/efeitos dos fármacos , Autofagia/genética , Cruzamentos Genéticos , Cicloeximida/farmacologia , Epistasia Genética/efeitos dos fármacos , Metanossulfonato de Etila , Regulação da Expressão Gênica de Plantas/efeitos dos fármacos , Teste de Complementação Genética , Leucina/análogos & derivados , Leucina/farmacologia , Luciferases/metabolismo , Mutação de Sentido Incorreto/genética , Fenótipo , Plantas Geneticamente Modificadas , Estabilidade Proteica/efeitos dos fármacos , Proteólise/efeitos dos fármacos , RNA Mensageiro/genética , RNA Mensageiro/metabolismo , Estresse Fisiológico/efeitos dos fármacos , Estresse Fisiológico/genética , Fatores de TempoRESUMO
In Arabidopsis, there are four homologs of class A1 heat shock factor (HSFA1) genes, which likely encode the master regulators of heat shock response (HSR). However, previous studies with double knockout (KO) mutants were unable to confirm this point probably due to functional redundancy. Here, we generated a quadruple KO (QK) and four triple KO mutants to dissect their functions. Our data show that members of the HSFA1 group not only play a pivotal role in HSR but also are involved in growth and development. Alterations in morphology and retardation in growth were observed in the quadruple but not in triple KO mutants. The basal and acquired thermotolerance capacity was dramatically decreased in the QK mutant but varied in triple KO mutants at different developmental stages. The transcriptomics profiles suggested that more than 65% of the heat stress (HS)-up-regulated genes were HSFA1 dependent. HSFA1s were also involved in the expression of several HS genes induced by H(2) O(2) , salt and mannitol, which is consistent with the increased sensitive phenotype of the QK mutant to the stress factors. In conclusion, the Arabidopsis HSFA1s function as the master regulators of HSR and participate as important components in other abiotic stress responses as well.
Assuntos
Proteínas de Arabidopsis/fisiologia , Arabidopsis/fisiologia , Proteínas de Ligação a DNA/fisiologia , Proteínas de Choque Térmico/fisiologia , Resposta ao Choque Térmico , Proteínas de Plantas/fisiologia , Fatores de Transcrição/fisiologia , Arabidopsis/genética , Arabidopsis/crescimento & desenvolvimento , Regulação da Expressão Gênica de Plantas , Técnicas de Inativação de Genes , Fatores de Transcrição de Choque Térmico , Mutação , Análise de Sequência com Séries de Oligonucleotídeos , RNA de Plantas/genética , Estresse FisiológicoRESUMO
Serpentine minerals with high levels of geologic chromium (Cr) and nickel (Ni) and non-serpentine farmlands polluted by irrigation water causing high anthropogenic Cr and Ni levels are both found in Taiwan. Elevated levels of Cr and Ni in these soils are a concern due to their potential to promote cancer mortality in humans. Bioaccessibility is a crucial factor determining the actual health risk via oral ingestion when children are exposed to metal-contaminated soils. Furthermore, the bioaccessibility of metals varies with the source, soil properties, and fractionation of metals in the soil. Therefore in this study, soil pH, total organic carbon (TOC), texture, and the total concentrations, fractionation, and bioaccessibility of Cr and Ni were analyzed and correlated for soils collected from serpentine mineral-containing deposits and contaminated non-serpentine farmlands. The low bioaccessibility and low mobility of Cr and Ni in serpentine soils suggested that incidental ingesting of soils posed a low health risk; however, the higher bioaccessibility and mobility of Ni in non-serpentine soils contaminated by electroplating wastewater could lead to potential risks for humans. Additionally, a significant difference in the bioaccessibility of Ni was observed between serpentine and non-serpentine soils, but this was not shown for Cr. Accordingly, a correlation analysis showed that Cr bioaccessibility was positively correlated with TOC, with no distinction between serpentine and non-serpentine soils. In contrast, TOC and the fractions of the sequential extraction procedure were significantly correlated with Ni bioaccessibility both in anthropogenically contaminated non-serpentine soils and in natural serpentine soils.