Can Nep1-like proteins form oligomers?

Cechin, Adelmo Luis; Sinigaglia, Marialva; Mombach, José C. M.; Echeverrigaray, Sérgio; Lemke, Ney; Cabrera, Odalys García; Pereira, Gonçalo Amarante Guimarães; Medrano, F.J.

doi:10.4161/psb.3.10.6643

Cited by 3 publications

(3 citation statements)

References 4 publications

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“…Previous reports raised the issue that NLPs may form oligomers in solution, especially dimers [23] , [31] . Additionally, it is well established that upon WBD progression, MpNep2 rather than MpNep1 accumulates in infected tissues, reaching its maximum concentration at the advanced necrotic stage [20] .…”

Section: Discussionmentioning

confidence: 99%

“…The oligomeric profile of MpNep2 has not been studied previously. Cechin and colleagues [31] discussed how NLPs could form oligomers based on sequence and structural similarities to the Cupin superfamily, a diverse group of proteins containing a conserved barrel domain known as cupa . MpNep2 protein was believed to act primarily in biotrophic mycelia as a monomer.…”

Section: Discussionmentioning

confidence: 99%

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Moniliophthora perniciosa Necrosis- and Ethylene-Inducing Protein 2 (MpNep2) as a Metastable Dimer in Solution: Structural and Functional Implications

et al. 2012

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Understanding how Nep-like proteins (NLPs) behave during the cell cycle and disease progression of plant pathogenic oomycetes, fungi and bacteria is crucial in light of compelling evidence that these proteins play a role in Witches` Broom Disease (WBD) of Theobroma cacao, one of the most important phytopathological problems to afflict the Southern Hemisphere. The crystal structure of MpNep2, a member of the NLP family and the causal agent of WBD, revealed the key elements for its activity. This protein has the ability to refold after heating and was believed to act as a monomer in solution, in contrast to the related homologs MpNep1 and NPP from the oomyceteous fungus Phytophthora parasitica. Here, we identify and characterize a metastable MpNep2 dimer upon over-expression in Escherichia coli using different biochemical and structural approaches. We found using ultra-fast liquid chromatography that the MpNep2 dimer can be dissociated by heating but not by dilution, oxidation or high ionic strength. Small-angle X-ray scattering revealed a possible tail-to-tail interaction between monomers, and nuclear magnetic resonance measurements identified perturbed residues involved in the putative interface of interaction. We also explored the ability of the MpNep2 monomer to refold after heating or chemical denaturation. We observed that MpNep2 has a low stability and cooperative fold that could be an explanation for its structure and activity recovery after stress. These results can provide new insights into the mechanism for MpNep2′s action in dicot plants during the progression of WBD and may open new avenues for the involvement of NLP- oligomeric species in phytopathological disorders.

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Section: Discussionmentioning

confidence: 99%

Section: Discussionmentioning

confidence: 99%

Moniliophthora perniciosa Necrosis- and Ethylene-Inducing Protein 2 (MpNep2) as a Metastable Dimer in Solution: Structural and Functional Implications

et al. 2012

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“…NEP1-like proteins (NLPs) occur in oomycetes, fungi, and bacteria and induce a hypersensitive-like response in a large number of dicotyledonous plants [2][3][4][5][6]. The 3D-structure of the NLPs remains to be determined experimentally, however recent bioinformatics studies have suggested that they belong to the Cupin superfamily [7,8]. NLPs seem to have a significant role in pathogenicity and these proteins may contribute to virulence since they are prevalent especially in plant pathogenic organisms (review in Ref.…”

Section: Introductionmentioning

confidence: 99%

Analysis of expression pathways alterations of Arabidopsis thaliana induced by a Necrosis- and Ethylene-inducing protein

Sinigaglia

Castro

Echeverrigaray

et al. 2009

Physica A: Statistical Mechanics and its Applications

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a b s t r a c tA major goal in post-genomic biology is the description of physiological functions in terms of gene pathway behavior. In this work, we present the first investigation of expression alterations in ninety-three pathways representing physiological functions of the plant A. thaliana induced by a P. parasitica elicitor (NLPpp) using microarray data publicly available at the AtGenExpress database. Using a novel statistical analysis developed to detect pathway alterations, we identified the gene pathways, hereby called groups of functionally associated genes (defined according to the TAIR/Gene Ontology and other databases), that are significantly altered in response to the elicitor. Instead of looking at individual gene responses, our analysis allowed a detailed characterization of the time ordering of pathways alterations in response to the NLPpp, their physiological implications and specificity. We also observed the activation of genes associated with vesicle trafficking and ROS production implying the initiation of the senescence of the wounded plant tissue.

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Use of iRNA in the post-transcriptional gene silencing of necrosis-inducing Phytophthora protein 1(NPP1) in Phytophthora cinnamomi

Pascoal-Ferreira,

Chahed,

Costa

et al. 2023

Mol Biol Rep

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Background Phytophthora cinnamomi is an Oomycetes associated with soil, this Oomycete is one of the most destructive species of Phytophthora, being responsible for the decline of more than 5000 ornamental, forest, or fruit plants. It can secrete a class of protein NPP1 (Phytophthora necrosis inducing protein 1), responsible for inducing necrosis in leaves and roots of plants, leading to their death. Objective This work will report the characterization of the Phytophthora cinnamomi NPP1 gene responsible for the infection of Castaneasativa roots and will characterize the mechanisms of interaction between Phytophthora cinnamomi and Castanea sativa, by gene silencing NPP1 from Phytophthora cinnamomi mediated by RNAi. Methods and results For silencing a part of the coding region of the NPP1 gene, was placed in the sense and antisense directions between an intron and ligated to the integrative vector pTH210. Cassette integration was confirmed by PCR and sequencing on the hygromycin-resistant Phytophthora cinnamomi transformants. Transformants obtained with the silenced gene was used to infect Castanea sativa. Conclusions Plants infected with these transformants showed a great reduction in disease symptoms, confirming iRNA as a potential alternative biological tool in the study of molecular factors, and in the control and management of Phytophthora cinnamomi.

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Can Nep1-like proteins form oligomers?

Cited by 3 publications

References 4 publications

Moniliophthora perniciosa Necrosis- and Ethylene-Inducing Protein 2 (MpNep2) as a Metastable Dimer in Solution: Structural and Functional Implications

Moniliophthora perniciosa Necrosis- and Ethylene-Inducing Protein 2 (MpNep2) as a Metastable Dimer in Solution: Structural and Functional Implications

Analysis of expression pathways alterations of Arabidopsis thaliana induced by a Necrosis- and Ethylene-inducing protein

Use of iRNA in the post-transcriptional gene silencing of necrosis-inducing Phytophthora protein 1(NPP1) in Phytophthora cinnamomi

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