RRC ID 56994
Author Endo H, Tanaka S, Adegawa S, Ichino F, Tabunoki H, Kikuta S, Sato R.
Title Extracellular loop structures in silkworm ABCC transporters determine their specificities for Bacillus thuringiensis Cry toxins.
Journal J Biol Chem
Abstract Bacillus thuringiensis Cry toxins are insecticidal proteins used widely for pest control. They are lethal to a restricted range of insects via specific interactions with insect receptors such as the ABC transporter subfamily members C2 (ABCC2) and C3 (ABCC3). However, it is still unclear how these different receptors contribute to insect susceptibility to Cry1A toxins. Here, we investigated the differences between the silkworm (Bombyx mori) ABCC2 (BmABCC2_S) and ABCC3 (BmABCC3) receptors in mediating Cry toxicity. Compared with BmABCC2_S, BmABCC3 exhibited 80- and 267-fold lower binding affinities to Cry1Aa and Cry1Ab, respectively, and these decreased affinities correlated well with the lower receptor activities of BmABCC3 for these Cry1A toxins. To identify the amino acid residues responsible for these differences, we constructed BmABCC3 variants containing a partial amino acid replacement with extracellular loops (ECLs) from BmABCC2_S. Replacing three amino acids from ECL 1 or 3 increased BmABCC3 activity toward Cry1Aa and enabled its activity toward Cry1Ab. Meanwhile, BmABCC2_S and BmABCC3 exhibited no receptor activities for Cry1Ca, Cry1Da, and Cry3Bb, correlating with markedly lower binding affinities for these Cry toxins. ABCC2 from a Cry1Ab-resistant B. mori strain (BmABCC2_R), which has a tyrosine insertion in ECL 2, displayed 93-fold lower binding affinity to Cry1Ab compared with BmABCC2_S but maintained high binding affinity to Cry1Aa. These results indicate that the Cry toxin-binding affinities of ABCC transporters are largely linked to the level of Cry susceptibility of ABCC-expressing cells and that the ABCC ECL structures determine the specificities to Cry toxins.
Volume 293(22)
Pages 8569-8577
Published 2018-6-1
DOI 10.1074/jbc.RA118.001761
PII RA118.001761
PMID 29666188
PMC PMC5986200
MeSH ATP-Binding Cassette Transporters / chemistry* ATP-Binding Cassette Transporters / metabolism* Animals Bacterial Proteins / chemistry Bacterial Proteins / metabolism* Bombyx / growth & development Bombyx / metabolism* Endotoxins / chemistry Endotoxins / metabolism* HEK293 Cells Hemolysin Proteins / chemistry Hemolysin Proteins / metabolism* Humans Insect Proteins / chemistry* Insect Proteins / metabolism* Insecticides / chemistry Insecticides / metabolism* Protein Conformation Substrate Specificity
IF 4.106
Times Cited 9