Cryptic endotoxic nature of <i>Bacillus thuringiensis</i> Cry1Ab insecticidal crystal protein

Vázquez-Padrón, Roberto I.; Riva, G; Agüero, Guillermin; Silva, Yussun; Pham, Si M.; Soberón, Mário; Bravo, Alejandra; Aı̈touche, Abdelouahab

doi:10.1016/j.febslet.2004.06.021

Cited by 18 publications

(8 citation statements)

References 30 publications

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“…The bactericidal character of the truncated versions cy ‐α Ctr and cy‐ α Ntr constructed here may point to possible proteolytic activation that it must undergo in vivo to expose its toxicity to some yet unknown target organism. The cryptic antibacterial action of the C‐terminal domain of Cry1Ab is similarly hidden when produced in contiguity with the N‐terminal insecticidal part of the toxin [22]. Sequestration of antibacterial activity of Cyt1Ca in a specific fold of the whole protein may provide a safety mechanism to protect the host bacterium.…”

Section: Discussionmentioning

confidence: 99%

Exposing cryptic antibacterial activity in Cyt1Ca from Bacillus thuringiensis israelensis by genetic manipulations

Itsko

Zaritsky

2007

FEBS Letters

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In an attempt to endow Cyt1Ca with Cyt1Aa-like antibacterial activity, both derived from Bacillus thuringiensis subsp. israelensis, two amino acids were replaced, E117V and N125A, so as to raise the hydrophobicity of the corresponding region, considered to be the membrane-active motif. The clones obtained included multiple repeats of VIEVLKSLLGIALA, corresponding to head-to-tail polymerization of the primer, translated in frame with Cyt1Ca. These versions of Cyt1Ca caused instant arrest in biomass growth and decreased viability upon expression in Escherichia coli. Multiple insertions of the non-mutated motif VIEELKSLLGINLA into the polypeptide were also lethal. To expose toxicity of the latter motif in the original Cyt1Ca, cyt1Ca was appropriately truncated.

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Section: Discussionmentioning

confidence: 99%

Exposing cryptic antibacterial activity in Cyt1Ca from Bacillus thuringiensis israelensis by genetic manipulations

Itsko

Zaritsky

2007

FEBS Letters

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“…Molecular biology experimental techniques allowed our group to study sequences relevant for biochemistry. 32,67 The experimental techniques used herein allowed us to isolate for the first time a possible ACO DNA partial sequence from Coffea arabica cv. caturra rojo.…”

Section: Comparison Of Different Molecular Descriptors Using Linear Cmentioning

confidence: 99%

2D‐RNA‐coupling numbers: A new computational chemistry approach to link secondary structure topology with biological function

et al. 2007

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Methods for prediction of proteins, DNA, or RNA function and mapping it onto sequence often rely on bioinformatics alignment approach instead of chemical structure. Consequently, it is interesting to develop computational chemistry approaches based on molecular descriptors. In this sense, many researchers used sequence-coupling numbers and our group extended them to 2D proteins representations. However, no coupling numbers have been reported for 2D-RNA topology graphs, which are highly branched and contain useful information. Here, we use a computational chemistry scheme: (a) transforming sequences into RNA secondary structures, (b) defining and calculating new 2D-RNA-coupling numbers, (c) seek a structure-function model, and (d) map biological function onto the folded RNA. We studied as example 1-aminocyclopropane-1-carboxylic acid (ACC) oxidases known as ACO, which control fruit ripening having importance for biotechnology industry. First, we calculated tau(k)(2D-RNA) values to a set of 90-folded RNAs, including 28 transcripts of ACO and control sequences. Afterwards, we compared the classification performance of 10 different classifiers implemented in the software WEKA. In particular, the logistic equation ACO = 23.8 . tau(1)(2D-RNA) + 41.4 predicts ACOs with 98.9%, 98.0%, and 97.8% of accuracy in training, leave-one-out and 10-fold cross-validation, respectively. Afterwards, with this equation we predict ACO function to a sequence isolated in this work from Coffea arabica (GenBank accession DQ218452). The tau(1)(2D-RNA) also favorably compare with other descriptors. This equation allows us to map the codification of ACO activity on different mRNA topology features. The present computational-chemistry approach is general and could be extended to connect RNA secondary structure topology to other functions.

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“…This result suggests that some other sequence stretch in the C‐terminal extension, such as block 6, also had a role in maintaining the structural stability of GST incorporated into the inclusions. The toxicity of the Cry1Ab C‐terminal extension against E. coli and Agrobacterium tumefaciens has been reported previously [18]. The toxicity is reduced or neutralized when the C‐terminal region is fused with domain III of the insecticidal N‐terminal region.…”

Section: Discussionmentioning

confidence: 71%

Novel strategy for protein production using a peptide tag derived from Bacillus thuringiensis Cry4Aa

et al. 2010

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Numerous proteins cannot be sufficiently prepared by ordinary recombinant DNA techniques because they are unstable or have deleterious effects on the host cell. One idea to prepare such proteins is to produce them as protein inclusions. Here we developed a novel system to effectively prepare proteins by using peptide tags derived from the insecticidal Cry toxin of a soil bacterium, Bacillus thuringiensis. Fusion with this peptide tag, designated 4AaCter, facilitates the formation of protein inclusions of glutathione S‐transferase in Escherichia coli without losing the enzyme activity. Application of 4AaCter to the production of syphilis antigens TpN15, TpN17 and TpN47 from Treponema pallidum yielded excellent results, including a dramatic increase in the production level, simplification of the product purification and high reactivity with syphilis antibody. The use of 4AaCter may provide an innovational strategy for the efficient production of proteins.

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Cryptic endotoxic nature of Bacillus thuringiensis Cry1Ab insecticidal crystal protein

Cited by 18 publications

References 30 publications

Exposing cryptic antibacterial activity in Cyt1Ca from Bacillus thuringiensis israelensis by genetic manipulations

Exposing cryptic antibacterial activity in Cyt1Ca from Bacillus thuringiensis israelensis by genetic manipulations

2D‐RNA‐coupling numbers: A new computational chemistry approach to link secondary structure topology with biological function

Novel strategy for protein production using a peptide tag derived from Bacillus thuringiensis Cry4Aa

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