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Arylphorin is a mitogen in the Heliothis virescens midgut cell secretome upon Cry1Ac intoxication

, , , , ,
1 Entomology and Plant Pathology, University of Tennessee, Knoxville, Tennessee, United States
2 Genome Science and Technology Graduate Program, University of Tennessee, Knoxville, Tennessee, United States
3 Southern Insect Management Research Unit, USDA-ARS, Stoneville, MS, USA
4 Forestry, Wildlife and Fisheries, University of Tennessee, Knoxville, Tennessee, United States
DOI
10.7287/peerj.preprints.2878v1
Published
Accepted
Subject Areas
Entomology, Microbiology, Histology
Keywords
arylphorin, Bacillus thuringiensis, Heliothis virescens, regeneration, stem cell, midgut, Cry1Ac
Copyright
© 2017 Castagnola et al.
Licence
This is an open access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, reproduction and adaptation in any medium and for any purpose provided that it is properly attributed. For attribution, the original author(s), title, publication source (PeerJ Preprints) and either DOI or URL of the article must be cited.
Cite this article
Castagnola A, Jackson J, Perera OP, Oppert C, Eda S, Jurat-Fuentes JL. 2017. Arylphorin is a mitogen in the Heliothis virescens midgut cell secretome upon Cry1Ac intoxication. PeerJ Preprints 5:e2878v1

Abstract

Insecticidal crystal (Cry) proteins produced by the bacterium Bacillus thuringiensis (Bt) target cells in the midgut epithelium of susceptible larvae. While the mode of action of Cry toxins has been extensively investigated, the midgut response to Cry intoxication and its regulation are not well characterized. In this work, we report the secreted proteome (secretome) of primary mature midgut cell cultures from Heliothis virescens larvae after exposure to Cry1Ac toxin compared to control buffer treatment. Biological activity of the Cry1Ac-induced secretome was monitored as higher proliferation and differentiation and an overall reduction in total cell mortality over time in primary H. virescens midgut stem cell cultures when compared to treatment with control buffer secretome. Differential proteomics identified 4 proteins with significant differences in abundance comparing Cry1Ac-treated and control secretomes. The most significant difference detected in the Cry1Ac secretome was an arylphorin protein not detected in the control secretome. Feeding of purified arylphorin to H. virescens larvae resulted in midgut hyperplasia and significantly reduced susceptibility to Cry1Ac toxin compared to controls. These data identify arylphorin as a protein with a putative relevant role in the midgut regeneration process in response to Cry1Ac intoxication.

Author Comment

This is a submission to PeerJ for review.

Supplemental Information

Proteins identified in primary midgut cell secretomes

DOI: 10.7287/peerj.preprints.2878v1/supp-1

Raw data used to prepare the figures in the paper

DOI: 10.7287/peerj.preprints.2878v1/supp-2

Additional Information

Competing Interests

The authors declare that they have no competing interests.

Author Contributions

Anais Castagnola conceived and designed the experiments, performed the experiments, analyzed the data, wrote the paper, prepared figures and/or tables, reviewed drafts of the paper.

Jerreme Jackson conceived and designed the experiments, performed the experiments, analyzed the data, wrote the paper, prepared figures and/or tables.

Omaththage P Perera conceived and designed the experiments, contributed reagents/materials/analysis tools, wrote the paper.

Cris Oppert conceived and designed the experiments, analyzed the data, contributed reagents/materials/analysis tools, wrote the paper.

Shigetoshi Eda conceived and designed the experiments, performed the experiments, analyzed the data, contributed reagents/materials/analysis tools, wrote the paper.

Juan Luis Jurat-Fuentes conceived and designed the experiments, analyzed the data, contributed reagents/materials/analysis tools, wrote the paper, prepared figures and/or tables, reviewed drafts of the paper.

Data Deposition

The following information was supplied regarding data availability:

The raw data has been supplied as a supplementary file.

Funding

This work was supported by award National Science Foundation Career Advancement Award number IOS-0718807. Partial funding was provided by Biotechnology Risk Assessment Grant Program competitive grant No. 2010-33522-21700 from the USDA National Institute of Food and Agriculture and a DuPont Young Professor Award from DuPont to JLJ-F. This work was also supported by the Postdoctoral Program for Excellence in Research and Teaching (PERT) at The University of Arizona Center for Insect Science, funding through the National Institute of Health training grant number 1K12 GM 00708, and the IRACDA Program, which supported A. Castagnola during the time of the writing of this manuscript. The funders had no role in study design, data collection and analysis, decision to publish, or preparation of the manuscript.

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Arylphorin is a mitogen in the Heliothis virescens midgut cell secretome upon Cry1Ac intoxication
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