Insect Biochemistry and Molecular Biology 28 (1998) 759–766 Isolation and identification of the cDNA encoding the pheromone biosynthesis activating neuropeptide and additional neuropeptides in the oriental tobacco budworm, Helicoverpa assulta (Lepidoptera: Noctuidae) 1 Man Yeon Choi a,* , Minoru Tanaka b , Hiroshi Kataoka b , Kyung Saeng Boo c , Sadahiro Tatsuki a a Department of Agricultural and Environmental Biology, Graduate School of Agricultural and Life Sciences, The University of Tokyo, Yayoi 1- 1-1, Bunkyo-ku, Tokyo 113, Japan b Department of Biotechnology, Graduate School of Agriculture and Life Sciences, The University of Tokyo, Yayoi 1-1-1, Bunkyo-ku, Tokyo 113, Japan c Division of Applied Biology and Chemistry, College of Agriculture and Life Sciences, Seoul National University, Suwon 441-744, South Korea Received 24 October 1997; received in revised form 12 June 1998; accepted 16 June 1998 Abstract The present study is concerned with cloning and characterizing Has-PBAN cDNA which is 756 nucleotides long, isolated from the brain and suboesophageal ganglion complex (Br–Sg) of Helicoverpa assulta adults. The 194-amino acid sequence deduced from this cDNA possessed the proteolytic endocleavage sites to generate multiple peptides. From the processing of the prepro-hormone, it can be predicted that the cDNA has a PBAN domain with 33 amino acids and four additional peptide domains: 24 amino acid-, 7 amino acid-, 18 amino acid- and 8 amino acid-long sequences, with FXPR(or K)L (X = G, T or S) amidated at their C-termini. The amino acid sequence of all five predicted peptides, including the PBAN, are identical to that of Helicoverpa zea (Raina, A.K., Jaffe, H., Kempe, T.G., Keim, P., Blacher, R.W., Fales, H.M., Riley, C.T., Klun, J.A., Ridgway, R.L., Hayes, D.K., 1989. Identifi- cation of a neuropeptide hormone that regulates sex pheromone production in female moths. Science 244, 796–798 and Ma, P.W.K., Knipple, D.C., Roelofs, W.L., 1994. Structural organization of the Helicoverpa zea gene encoding the precursor protein for phero- mone biosynthesis-activating neuropeptide and other neuropeptides. Proc. Natl. Acad. Sci., U.S.A. 91, 506–510). A single mRNA species corresponding to the size of Has-PBAN cDNA was detected from the Br–Sg of 1–3-day old female and male adults, and their expression was also at a similar level. Pheromone production was induced upon injection of female or male Br–Sg extracts or synthetic PBAN into the haemocoel of decapitated 1–3-day old female adults during the photophase when they are not supposed to produce pheromone. From these results, H. assulta adult females seem to use their own PBAN for regulating sex pheromone biosynthesis. Functions of the four other peptides ending with FXPR(or K)L in the Has-PBAN cDNA and of the male PBAN remain to be elucidated.  1998 Elsevier Science Ltd. All rights reserved. Keywords: Helicoverpa assulta; Has-PBAN; cDNA; Regulation of sex pheromone production; Z9-16:Ald 1. Introduction Most female moths use sex pheromones for chemical communication such as attracting and mating with their male counterparts. Sex pheromone biosynthesis is usu- * Corresponding author. Tel.: + 81-3-3812-2111 (ext. 5093); Fax: + 81-3-3815-5851; e-mail: aa57401@hongo.ecc.u-tokyo.ac.jp 1 The cDNA sequence of this paper has been deposited in the Gen- Bank data base (Accession No. U96761). 0965-1748/98/$19.00  1998 Elsevier Science Ltd. All rights reserved. PII:S0965-1748(98)00065-4 ally regulated by hormonal and/or neural factors. A pher- omonotropic factor such as pheromone biosynthesis acti- vating neuropeptide (PBAN) is synthesized in the suboesophageal ganglion and released into the haemo- lymph to stimulate pheromone biosynthesis in the phero- mone gland of female moths (Raina and Klun, 1984). The primary structure of PBAN has been reported from three moth species, Hez-PBAN from Helicoverpa zea (Raina et al., 1989), Bom-PBAN I and II from Bom- byx mori (Kitamura et al., 1989, 1990) and Lyd-PBAN