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Disruption of Ha_BtR alters binding of Bacillus thuringiensis delta-endotoxin Cry1Ac to midgut BBMVs of Helicoverpa armigera.
J Invertebr Pathol. 2008 Jan; 97(1):27-32.JI

Abstract

Disruption of the Ha_BtR (a cadherin gene) is genetically linked to resistance to Cry1Ac delta-endotoxin of Bacillus thuringiensis in the GYBT strain of Helicoverpa armigera. Brush border membrane vesicles (BBMVs) prepared from midguts of both the Cry1Ac-resistant GYBT strain (homozygous for a deletion knockout of Ha_BtR) and the susceptible GY strain (homozygous for the wild type of Ha_BtR) possessed saturable and specific binding ability to (125)I-Cry1Ac. The binding constant (K(d)) of the GY strain was significantly lower than that of the resistant GYBT strain, whereas their binding site concentrations (B(max)) were similar. When midgut BBMVs were reacted directly with streptavidin conjugated to horseradish peroxidase, the GY strain had very clear 120- and 85-kDa protein bands, which indicated that the 120- and 85-kDa bands are endogenous biotin-containing proteins. However, the GYBT strain almost completely lost these two biotin-containing proteins. Ligand blotting with biotinylated Cry1Ac toxin showed midgut BBMVs of the GY strain contain five protein bands of 210-, 190-, 150-, 120-, and 85-kDa, respectively, while BBMVs of the GYBT strain contain only two protein bands of 150- and 120-kDa. 120-kDa bands may consist of two proteins with coincidentally the same molecular weight (putatively, an APN and a biotin-containing protein). Our results showed that the binding pattern of Cry1Ac to midgut BBMVs of H. armigera was altered quantitatively and qualitatively by knockout of Ha_BtR. There are multiple Cry1Ac-binding proteins in the midgut of susceptible H. armigera, but only the Ha_BtR can be considered as a putative functional receptor of Cry1Ac. Possible involvement of other receptor proteins in the intoxication process in vivo could not be excluded.

Authors+Show Affiliations

Department of Entomology, College of Plant Protection, Nanjing Agricultural University, Nanjing 210095, China.No affiliation info available

Pub Type(s)

Journal Article
Research Support, Non-U.S. Gov't

Language

eng

PubMed ID

17681529

Citation

Xu, Xinjun, and Yidong Wu. "Disruption of Ha_BtR Alters Binding of Bacillus Thuringiensis Delta-endotoxin Cry1Ac to Midgut BBMVs of Helicoverpa Armigera." Journal of Invertebrate Pathology, vol. 97, no. 1, 2008, pp. 27-32.
Xu X, Wu Y. Disruption of Ha_BtR alters binding of Bacillus thuringiensis delta-endotoxin Cry1Ac to midgut BBMVs of Helicoverpa armigera. J Invertebr Pathol. 2008;97(1):27-32.
Xu, X., & Wu, Y. (2008). Disruption of Ha_BtR alters binding of Bacillus thuringiensis delta-endotoxin Cry1Ac to midgut BBMVs of Helicoverpa armigera. Journal of Invertebrate Pathology, 97(1), 27-32.
Xu X, Wu Y. Disruption of Ha_BtR Alters Binding of Bacillus Thuringiensis Delta-endotoxin Cry1Ac to Midgut BBMVs of Helicoverpa Armigera. J Invertebr Pathol. 2008;97(1):27-32. PubMed PMID: 17681529.
* Article titles in AMA citation format should be in sentence-case
TY - JOUR T1 - Disruption of Ha_BtR alters binding of Bacillus thuringiensis delta-endotoxin Cry1Ac to midgut BBMVs of Helicoverpa armigera. AU - Xu,Xinjun, AU - Wu,Yidong, Y1 - 2007/07/04/ PY - 2007/01/18/received PY - 2007/04/26/revised PY - 2007/04/27/accepted PY - 2007/8/8/pubmed PY - 2008/2/21/medline PY - 2007/8/8/entrez SP - 27 EP - 32 JF - Journal of invertebrate pathology JO - J Invertebr Pathol VL - 97 IS - 1 N2 - Disruption of the Ha_BtR (a cadherin gene) is genetically linked to resistance to Cry1Ac delta-endotoxin of Bacillus thuringiensis in the GYBT strain of Helicoverpa armigera. Brush border membrane vesicles (BBMVs) prepared from midguts of both the Cry1Ac-resistant GYBT strain (homozygous for a deletion knockout of Ha_BtR) and the susceptible GY strain (homozygous for the wild type of Ha_BtR) possessed saturable and specific binding ability to (125)I-Cry1Ac. The binding constant (K(d)) of the GY strain was significantly lower than that of the resistant GYBT strain, whereas their binding site concentrations (B(max)) were similar. When midgut BBMVs were reacted directly with streptavidin conjugated to horseradish peroxidase, the GY strain had very clear 120- and 85-kDa protein bands, which indicated that the 120- and 85-kDa bands are endogenous biotin-containing proteins. However, the GYBT strain almost completely lost these two biotin-containing proteins. Ligand blotting with biotinylated Cry1Ac toxin showed midgut BBMVs of the GY strain contain five protein bands of 210-, 190-, 150-, 120-, and 85-kDa, respectively, while BBMVs of the GYBT strain contain only two protein bands of 150- and 120-kDa. 120-kDa bands may consist of two proteins with coincidentally the same molecular weight (putatively, an APN and a biotin-containing protein). Our results showed that the binding pattern of Cry1Ac to midgut BBMVs of H. armigera was altered quantitatively and qualitatively by knockout of Ha_BtR. There are multiple Cry1Ac-binding proteins in the midgut of susceptible H. armigera, but only the Ha_BtR can be considered as a putative functional receptor of Cry1Ac. Possible involvement of other receptor proteins in the intoxication process in vivo could not be excluded. SN - 0022-2011 UR - https://www.unboundmedicine.com/medline/citation/17681529/Disruption_of_Ha_BtR_alters_binding_of_Bacillus_thuringiensis_delta_endotoxin_Cry1Ac_to_midgut_BBMVs_of_Helicoverpa_armigera_ L2 - https://linkinghub.elsevier.com/retrieve/pii/S0022-2011(07)00142-5 DB - PRIME DP - Unbound Medicine ER -