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The Journal of Immunology, 2007, 179: 6943-6951.
Copyright © 2007 by The American Association of Immunologists, Inc.
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Noduler, A Novel Immune Up-Regulated Protein Mediates Nodulation Response in Insects1

Archana S. Gandhe, Serene H. John and Javaregowda Nagaraju2

Centre of Excellence for Genetics and Genomics of Silkmoths, Laboratory of Molecular Genetics, Centre for DNA Fingerprinting and Diagnostics, ECIL Road, Nacharam, Hyderabad, India

Insect immune system comprises of both humoral and cellular defenses. Nodulation is one of the major, yet very poorly understood cellular responses against microbial infections in insects. Through screening for novel immune genes from an Indian saturniid silkmoth Antheraea mylitta, we identified a protein up-regulated in hemolymph within minutes upon bacterial challenge. We have shown here, for first time, the involvement of this novel protein in mediating nodulation response against bacteria and hence designated it as Noduler. Noduler possessed a characteristic reeler domain found in several extracellular matrix vertebrate proteins. Noduler was shown in vitro to bind a wide range of bacteria, yeast, and also insect hemocytes. Furthermore, Noduler specifically bound LPS, lipotechoic acid, and β-1, 3 glucan components of microbial cell walls. RNA-interference mediated knock-down of the Noduler resulted in significant reduction in the number of nodules and consequent increase in bacterial load in larval hemolymph. The results suggest that the Noduler is widely conserved and is involved in very early clearance of bacteria by forming nodules of hemocytes and bacterial complexes in insects. The results would promote further studies for understanding of the crucial but hitherto overlooked nodulation mechanism in insects and also provide cues for the study of similar mammalian proteins whose function is not understood.

The costs of publication of this article were defrayed in part by the payment of page charges. This article must therefore be hereby marked advertisement in accordance with 18 U.S.C. Section 1734 solely to indicate this fact.

1 A.S.G. is a recipient of research fellowship from University Grants Commission of India and this work is funded by Department of Biotechnology, Government of India under Centre of Excellence for Genetics and Genomics of Silkmoths Grant (to J.N.).

2 Address correspondence and reprint requests to Dr. J. Nagaraju, Laboratory of Molecular Genetics, Centre for DNA Fingerprinting and Diagnostics, ECIL Road, Nacharam, Hyderabad, India. E-mail address: jnagaraju{at}cdfd.org.in

3 Abbreviations used in this paper: RNAi, RNA interference; dsRNA, double stranded RNA; PTU, phenylthiourea; IBS, insect buffer saline; PGN, peptidoglycan; LTA, lipotechoic acid; GFP, green fluorescent protein.






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