Characterization of the Bombyx mori Cecropin A1 promoter regulated by IMD pathway

Cecropin A1 (CecA1) promoter from Bombyx mori was cloned and characterized to provide insight into the transcriptional control of this antimicrobial peptide gene upon immune challenges. Reporter gene assays demonstrated that both Escherichia coli and lipopolysaccharide could induce expression in BmE cells but B. bombyseptieus or peptidoglycan failed, and the induction pattern of the reporter gene was coincident with the endogenous CecA1. Analysis of deletion and mutation constructs revealed that the regulatory region was the κB motif located between ‐176 and ‐166, and no other predicted elements on CecA1 promoter affected its inducibility. Insertion of additional κB motifs increased the activity of CecA1 promoter. Furthermore, binding of Relish to κB motif was confirmed by electrophoretic mobility shift assay. These findings indicate the regulatory mechanism of CecA1 expression in IMD pathway and suggest an approach of engineering antimicrobial peptide promoter with enhanced activities that may lead to broad applications.

[1]  Q. Guo,et al.  Transgene-based, female-specific lethality system for genetic sexing of the silkworm, Bombyx mori , 2013, Proceedings of the National Academy of Sciences.

[2]  Q. Xia,et al.  An optimized sericin-1 expression system for mass-producing recombinant proteins in the middle silk glands of transgenic silkworms , 2013, Transgenic Research.

[3]  Q. Xia,et al.  Resistance to BmNPV via Overexpression of an Exogenous Gene Controlled by an Inducible Promoter and Enhancer in Transgenic Silkworm, Bombyx mori , 2012, PloS one.

[4]  Q. Xia,et al.  Genome-Wide Identification and Immune Response Analysis of Serine Protease Inhibitor Genes in the Silkworm, Bombyx mori , 2012, PloS one.

[5]  S. Goodbourn,et al.  Regulation of transcription of the Aedes albopictus cecropin A1 gene: A role for p38 mitogen-activated protein kinase. , 2011, Insect biochemistry and molecular biology.

[6]  M. Yamakawa,et al.  Regulation of the innate immune responses in the silkworm, Bombyx mori , 2011 .

[7]  Q. Xia,et al.  Functional Divergence among Silkworm Antimicrobial Peptide Paralogs by the Activities of Recombinant Proteins and the Induced Expression Profiles , 2011, PloS one.

[8]  Y. Ip,et al.  Heterodimers of NF-κB transcription factors DIF and Relish regulate antimicrobial peptide genes in Drosophila , 2010, Proceedings of the National Academy of Sciences.

[9]  Cheng Lu,et al.  Establishment and characterization of two embryonic cell lines of Bombyx mori , 2007, In Vitro Cellular & Developmental Biology - Animal.

[10]  Y. Nakahara,et al.  A transgenic silkworm expressing the immune-inducible cecropin B-GFP reporter gene. , 2006, Insect biochemistry and molecular biology.

[11]  Q. Xia,et al.  Structures, regulatory regions, and inductive expression patterns of antimicrobial peptide genes in the silkworm Bombyx mori. , 2006, Genomics.

[12]  Masafumi Yamamoto,et al.  A novel Rel protein and shortened isoform that differentially regulate antibacterial peptide genes in the silkworm Bombyx mori. , 2005, Biochimica et biophysica acta.

[13]  M. Levine,et al.  Immunity regulatory DNAs share common organizational features in Drosophila. , 2004, Molecular cell.

[14]  G. Rubin,et al.  The Toll and Imd pathways are the major regulators of the immune response in Drosophila , 2002, The EMBO journal.

[15]  M. Belvin,et al.  A genome-wide analysis of immune responses in Drosophila , 2001, Proceedings of the National Academy of Sciences of the United States of America.

[16]  K. Taniai,et al.  A Novel Lipopolysaccharide Response Element in the Bombyx mori Cecropin B Promoter* , 2000, The Journal of Biological Chemistry.

[17]  Han Zs,et al.  Interaction and Specificity of Rel-related Proteins in Regulating Drosophila Immunity Gene Expression , 1999 .

[18]  Y. Ip,et al.  Interaction and Specificity of Rel-related Proteins in Regulating Drosophila Immunity Gene Expression* , 1999, The Journal of Biological Chemistry.

[19]  H. Tanaka,et al.  Immune proteins and their gene expression in the silkworm, Bombyx mori. , 1999, Developmental and comparative immunology.

[20]  U. Petersen,et al.  Adjacent GATA and κB-like Motifs Regulate the Expression of a Drosophila Immune Gene , 1997 .

[21]  M. Yamakawa,et al.  Moricin, a Novel Type of Antibacterial Peptide Isolated from the Silkworm, Bombyx mori(*) , 1995, The Journal of Biological Chemistry.

[22]  Q. Xia,et al.  Identification and analysis of Toll-related genes in the domesticated silkworm, Bombyx mori. , 2008, Developmental and comparative immunology.