Cry toxin specificities of insect ABCC transporters closely related to lepidopteran ABCC2 transporters

Endo, Haruka; Tanaka, Shiho; Imamura, Kazuhiro; Adegawa, Satomi; Kikuta, Shingo; Satō, Ryōichi

doi:10.1016/j.peptides.2017.04.003

Cited by 33 publications

(54 citation statements)

References 34 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…It is still not known how ABCA2 is involved in the susceptibility of larvae to Cry2A toxins. ABCA2 is a membrane protein and the ABC transporter ABCC2 is a functional Cry1A toxin receptor [5,16,18,24,25]. Therefore, we examined whether BmABCA2 transiently expressed in HEK293T cells ( Figure 5) could function as a Cry2Ab receptor and whether HEK293T cells would swell like the columnar cells in a Cry toxin-intoxicated midgut.…”

Section: Discussionmentioning

confidence: 99%

“…HEK293T cells were cultured and transfected, as described previously [5]. The HEK293T cells were cultured on the cover glass in a 6-well plate (Truesline; Nippon Genetics, Tokyo, Japan).…”

Section: Expression Of Bmabca2 and Bmabcc2 In Hek293t Cells And Cell mentioning

confidence: 99%

“…This interaction drives the toxins to insert partial structures into the membrane, forming ion channels [4]. A cation influx triggers the influx of water [5], resulting in cell swelling and lysis [2,6]. Given their strong toxicity in specific species and inability to harm the environment and human health, Cry toxins are used widely in pest control [7].…”

Section: Introductionmentioning

confidence: 99%

See 2 more Smart Citations

ATP-Binding Cassette Subfamily a Member 2 Is a Functional Receptor for Bacillus thuringiensis Cry2A Toxins in Bombyx mori, But Not for Cry1A, Cry1C, Cry1D, Cry1F, or Cry9A Toxins

Miyamoto

Takasu

et al. 2020

Toxins

Self Cite

View full text Add to dashboard Cite

Cry toxins are insecticidal proteins produced by Bacillus thuringiensis (Bt). They are used commercially to control insect pests since they are very active in specific insects and are harmless to the environment and human health. The gene encoding ATP-binding cassette subfamily A member 2 (ABCA2) was identified in an analysis of Cry2A toxin resistance genes. However, we do not have direct evidence for the role of ABCA2 for Cry2A toxins or why Cry2A toxin resistance does not cross to other Cry toxins. Therefore, we performed two experiments. First, we edited the ABCA2 sequence in Bombyx mori using transcription activator-like effector-nucleases (TALENs) and confirmed the susceptibility-determining ability in a diet overlay bioassay. Strains with C-terminal half-deleted BmABCA2 showed strong and specific resistance to Cry2A toxins; even strains carrying a deletion of 1 to 3 amino acids showed resistance. However, the C-terminal half-deleted strains did not show cross-resistance to other toxins. Second, we conducted a cell swelling assay and confirmed the specific ability of BmABCA2 to Cry2A toxins in HEK239T cells. Those demonstrated that BmABCA2 is a functional receptor for Cry2A toxins and that BmABCA2 deficiency-dependent Cry2A resistance does not confer cross-resistance to Cry1A, Cry1Ca, Cry1Da, Cry1Fa or Cry9Aa toxins.

show abstract

Section: Discussionmentioning

confidence: 99%

“…HEK293T cells were cultured and transfected, as described previously [5]. The HEK293T cells were cultured on the cover glass in a 6-well plate (Truesline; Nippon Genetics, Tokyo, Japan).…”

Section: Expression Of Bmabca2 and Bmabcc2 In Hek293t Cells And Cell mentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

See 1 more Smart Citation

ATP-Binding Cassette Subfamily a Member 2 Is a Functional Receptor for Bacillus thuringiensis Cry2A Toxins in Bombyx mori, But Not for Cry1A, Cry1C, Cry1D, Cry1F, or Cry9A Toxins

Miyamoto

Takasu

et al. 2020

Toxins

Self Cite

View full text Add to dashboard Cite

show abstract

“…Generating PxABCC2 mutation in the G88 wild-type strain led to a slightly higher resistance to Cry1Ac protoxin, yet PxABCC3 mutations caused higher susceptibility. Similarly, cultured cells of B. mori and S. exigua exhibit higher Cry1Aa susceptibility when expressing ABCC2 than ABCC3 [24,33]. Contribution of ABCC2 in determining toxicity of Cry1Ac may therefore be more significant than that of ABCC3.…”

Section: Plos Pathogensmentioning

confidence: 99%

“…The receptor functionality of ABCC2 for Cry1A toxins has been well characterized using heterologous expression in cultured cell lines, manipulating mRNA abundance with RNA interference and with transgenic Drosophila [14,[18][19][20][21][22]. ABC transporter subfamily C3 (ABCC3), a paralog of ABCC2, has also been reported to mediate Cry1A toxicity [14,[22][23][24]. Recently, Wang et al [25] reported mutations in ABCC2 plus ABCC3 were required to achieve highlevel Cry1Ac resistance in Helicoverpa armigera, suggesting functional redundancy as toxin receptors.…”

Section: Introductionmentioning

confidence: 99%

Resistance to Bacillus thuringiensis Cry1Ac toxin requires mutations in two Plutella xylostella ATP-binding cassette transporter paralogs

Xiang

et al. 2020

PLoS Pathog

View full text Add to dashboard Cite

The diamondback moth, Plutella xylostella, is a cosmopolitan pest and the first species to develop field resistance to toxins from the gram-positive bacterium Bacillus thuringiensis (Bt). Although previous work has suggested that mutations of ATP-binding cassette transporter subfamily C2 (ABCC2) or C3 (ABCC3) genes can confer Cry1Ac resistance, here we reveal that P. xylostella requires combined mutations in both PxABCC2 and PxABCC3 to achieve high-level Cry1Ac resistance, rather than simply a mutation of either gene. We identified natural mutations of PxABCC2 and PxABCC3 that concurrently occurred in a Cry1Acresistant strain (Cry1S1000) of P. xylostella, with a mutation (R A2) causing the mis-splicing of PxABCC2 and another mutation (R A3) leading to the premature termination of PxABCC3. Genetic linkage analysis showed that R A2 and R A3 were tightly linked to Cry1Ac resistance. Introgression of R A2 and R A3 enabled a susceptible strain (G88) of P. xylostella to obtain high resistance to Cry1Ac, confirming that these genes confer resistance. To further support the role of PxABCC2 and PxABCC3 in Cry1Ac resistance, frameshift mutations were introduced into PxABCC2 and PxABCC3 singly and in combination in the G88 strain with CRISPR/Cas9 mediated mutagenesis. Bioassays of CRISPR-based mutant strains, plus genetic complementation tests, demonstrated that the deletion of PxABCC2 or PxABCC3 alone provided < 4-fold tolerance to Cry1Ac, while disruption of both genes together conferred >8,000-fold resistance to Cry1Ac, suggesting the redundant/complementary roles of PxABCC2 and PxABCC3. This work advances our understanding of Bt resistance in P. xylostella by demonstrating mutations within both PxABCC2 and PxABCC3 genes are required for high-level Cry1Ac resistance.

show abstract

Silencing of an ABC transporter, but not a cadherin, decreases the susceptibility of Colorado potato beetle larvae to Bacillus thuringiensis ssp. tenebrionis Cry3Aa toxin

Güney

Cedden

Hänniger

et al. 2021

Arch Insect Biochem Physiol

View full text Add to dashboard Cite

The Colorado potato beetle, Leptinotarsa decemlineata (Coleoptera: Chrysomelidae), is a major pest of potato plants worldwide and is notorious for its ability to develop resistance to insecticides. Cry3 toxins synthesized by Bacillus thuringiensis ssp. tenebrionis have been used successfully to manage this pest. Resistance to Cry toxins is a concerning problem for many insect pests; therefore, it is important to determine the mechanisms by which insects acquire resistance to these toxins. Cadherin‐like and ABC transporter proteins have been implicated in the mode of action of Cry toxins as mutations in these genes render lepidopterans resistant to them; however, clear consensus does not exist on whether these proteins also play a role in Cry3 toxin activity and/or development of resistance in coleopterans. In the current study, we identified the L. decemlineata orthologues of the cadherin (LdCAD) and the ABCB transporter (LdABCB1) that have been implicated in the mode of action of Cry toxins in other coleopterans. Suppression of LdABCB1 via RNA interference reduced toxin‐related larval mortality, whereas partial silencing of LdCAD did not. Our results suggest that the ABCB is involved in the mode of action of Cry3Aa toxins; however, no evidence was found to support the role of cadherin as a receptor of Cry3Aa in L. decemlineata.

show abstract

Cry toxin specificities of insect ABCC transporters closely related to lepidopteran ABCC2 transporters

Cited by 33 publications

References 34 publications

ATP-Binding Cassette Subfamily a Member 2 Is a Functional Receptor for Bacillus thuringiensis Cry2A Toxins in Bombyx mori, But Not for Cry1A, Cry1C, Cry1D, Cry1F, or Cry9A Toxins

ATP-Binding Cassette Subfamily a Member 2 Is a Functional Receptor for Bacillus thuringiensis Cry2A Toxins in Bombyx mori, But Not for Cry1A, Cry1C, Cry1D, Cry1F, or Cry9A Toxins

Resistance to Bacillus thuringiensis Cry1Ac toxin requires mutations in two Plutella xylostella ATP-binding cassette transporter paralogs

Silencing of an ABC transporter, but not a cadherin, decreases the susceptibility of Colorado potato beetle larvae to Bacillus thuringiensis ssp. tenebrionis Cry3Aa toxin

Contact Info

Product

Resources

About