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1.
BMC Plant Biol ; 15: 198, 2015 08 14.
Artigo em Inglês | MEDLINE | ID: mdl-26268941

RESUMO

BACKGROUND: In Eucalyptus genus, studies on genome composition and transposable elements (TEs) are particularly scarce. Nearly half of the recently released Eucalyptus grandis genome is composed by retrotransposons and this data provides an important opportunity to understand TE dynamics in Eucalyptus genome and transcriptome. RESULTS: We characterized nine families of transcriptionally active LTR retrotransposons from Copia and Gypsy superfamilies in Eucalyptus grandis genome and we depicted genomic distribution and copy number in two Eucalyptus species. We also evaluated genomic polymorphism and transcriptional profile in three organs of five Eucalyptus species. We observed contrasting genomic and transcriptional behavior in the same family among different species. RLC_egMax_1 was the most prevalent family and RLC_egAngela_1 was the family with the lowest copy number. Most families of both superfamilies have their insertions occurring <3 million years, except one Copia family, RLC_egBianca_1. Protein theoretical models suggest different properties between Copia and Gypsy domains. IRAP and REMAP markers suggested genomic polymorphisms among Eucalyptus species. Using EST analysis and qRT-PCRs, we observed transcriptional activity in several tissues and in all evaluated species. In some families, osmotic stress increases transcript values. CONCLUSION: Our strategy was successful in isolating transcriptionally active retrotransposons in Eucalyptus, and each family has a particular genomic and transcriptional pattern. Overall, our results show that retrotransposon activity have differentially affected genome and transcriptome among Eucalyptus species.


Assuntos
Eucalyptus/genética , Genoma de Planta , Proteínas de Plantas/genética , Retroelementos , Transcriptoma , Eucalyptus/metabolismo , Dados de Sequência Molecular , Mutagênese Insercional , Filogenia , Proteínas de Plantas/química , Proteínas de Plantas/metabolismo , Estrutura Secundária de Proteína , Análise de Sequência de DNA , Sequências Repetidas Terminais
2.
Biomed Res Int ; 2014: 341270, 2014.
Artigo em Inglês | MEDLINE | ID: mdl-24696848

RESUMO

This paper shows the results of quercitrin effects on the structure and biological activity of secretory phospholipase (sPLA2) from Crotalus durissus terrificus, which is the main toxin involved in the pharmacological effects of this snake venom. According to our mass spectrometry and circular dichroism results, quercetin was able to promote a chemical modification of some amino acid residues and modify the secondary structure of C. d. terrificus sPLA2. Moreover, molecular docking studies showed that quercitrin can establish chemical interactions with some of the crucial amino acid residues involved in the enzymatic activity of the sPLA2, indicating that this flavonoid could also physically impair substrate molecule access to the catalytic site of the toxin. Additionally, in vitro and in vivo assays showed that the quercitrin strongly diminished the catalytic activity of the protein, altered its Vmax and Km values, and presented a more potent inhibition of essential pharmacological activities in the C. d. terrificus sPLA2, such as its myotoxicity and edematogenic effect, in comparison to quercetin. Thus, we concluded that the rhamnose group found in quercitrin is most likely essential to the antivenom activities of this flavonoid against C. d. terrificus sPLA2.


Assuntos
Venenos de Crotalídeos/toxicidade , Crotalus/metabolismo , Edema/patologia , Células Musculares/patologia , Fosfolipases A2 Secretórias/toxicidade , Quercetina/análogos & derivados , Animais , Dicroísmo Circular , Venenos de Crotalídeos/química , Venenos de Crotalídeos/isolamento & purificação , Ensaios Enzimáticos , Glicosilação/efeitos dos fármacos , Masculino , Camundongos , Simulação de Acoplamento Molecular , Células Musculares/efeitos dos fármacos , Fosfolipases A2 Secretórias/química , Fosfolipases A2 Secretórias/isolamento & purificação , Quercetina/química , Quercetina/farmacologia
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