Characterization and baculovirus-directed expression of a myosuppressin encoding cDNA from the true armyworm, Pseudaletia unipuncta

• Published in: Peptides (New York, N.Y. : 1980) N.Y. : 1980), 2002-04, Vol.23 (4), p.747-756
• Main Authors: Lee, E., Lange, A., Orchard, I., Fusé, M., Tobe, S.S., Bendena, W.G., Donly, B.C.
• Format: Article
• Language: English
• Subjects: Amino Acid Sequence; Animals; Baculoviridae - genetics; Baculoviridae - metabolism; Baculovirus; Base Sequence; DNA, Complementary; FMRFamide-related peptide; Hemolymph - chemistry; In Situ Hybridization; Insect; Insect Proteins - chemistry; Insect Proteins - genetics; Insect Proteins - metabolism; Molecular Sequence Data; Moths - genetics; Moths - metabolism; Moths - virology; Nerve Tissue - cytology; Nerve Tissue - metabolism; Neuropeptide; Neuropeptides - blood; Neuropeptides - chemistry; Neuropeptides - genetics; Noctuidae; Pseudaletia unipuncta; Sequence Alignment
• ISSN: 0196-9781; 1873-5169
• DOI: 10.1016/S0196-9781(01)00668-4

Insect myosuppressins are a highly conserved sub-family of peptides which are primarily characterized by the ability to suppress contraction of visceral muscles in a variety of insect species. We have isolated a cDNA from the true armyworm, Pseudaletia unipuncta, that encodes a prohormone containing a peptide identical to ManducaFLRFamide. We have shown that this myosuppressin gene appears to be expressed in late larval and adult insects. In Manduca sexta, a number of extended-FLRFamide peptides have previously been purified including ManducaFLRFamide, F7D (DPSFLRFamide), F7G (GNSFLRFamide) and two larger peptides F24 and F39 that contain the shorter ManducaFLRFamide sequence at their C-terminus. Comparison with the true armyworm prepropeptide characterized here identifies F24 and F39 as partially processed products from the same precursor. Expression in the true armyworm was shown by in situ hybridization to occur in over 150 cells throughout the adult brain and nerve cord, and also to occur in both open and closed endocrine type cells of the gut. Overexpression of the P. unipuncta FLRFamide cDNA from a baculovirus vector in cabbage looper caterpillars was used to assess the potential for myosuppressin expression as a means of enhancing virus efficacy. Viral expression of the armyworm prohormone cDNA resulted in raised levels of RFamide-like products in the hemolymph of infected insects, but the products were found to be chemically distinguishable from authentic mature peptide and probably represent partially processed hormone.