Identification of the gene encoding bursicon, an insect neuropeptide responsible for cuticle sclerotization and wing spreading.
To accommodate growth, insects must periodically replace their exoskeletons. After shedding the old cuticle, the new soft cuticle must sclerotize. Sclerotization has long been known to be controlled by the neuropeptide hormone bursicon, but its large size of 30 kDa has frustrated attempts to determine its sequence and structure. Using partial sequences obtained from purified cockroach bursicon, we identified the Drosophila melanogaster gene CG13419 as a candidate bursicon gene. CG13419 encodes a peptide with a predicted final molecular weight of 15 kDa, which likely functions as a dimer. This predicted bursicon protein belongs to the cystine knot family, which includes vertebrate transforming growth factor-beta (TGF-beta) and glycoprotein hormones. Point mutations in the bursicon gene cause defects in cuticle sclerotization and wing expansion behavior. Bioassays show that these mutants have decreased bursicon bioactivity. In situ hybridization and immunocytochemistry revealed that bursicon is co-expressed with crustacean cardioactive peptide (CCAP). Transgenic flies that lack CCAP neurons also lacked bursicon bioactivity. Our results indicate that CG13419 encodes bursicon, the last of the classic set of insect developmental hormones. It is the first member of the cystine knot family to have a defined function in invertebrates. Mutants show that the spectrum of bursicon actions is broader than formerly demonstrated.
Dewey
Elizabeth M
EM
Department of Biological Sciences, Vanderbilt University, Nashville, TN 37235, USA.
McNabb
Susan L
SL
Ewer
John
J
Kuo
Gloria R
GR
Takanishi
Christina L
CL
Truman
James W
JW
Honegger
Hans-Willi
HW
D000595:Amino Acid Sequence; D000818:Animals; D001681:Biological Assay; D017931:DNA Primers; D004331:Drosophila melanogaster / Q000235:genetics*; D007150:Immunohistochemistry; D017403:In Situ Hybridization; D007447:Invertebrate Hormones / Q000235:genetics* / Q000378:metabolism; D008969:Molecular Sequence Data; D018978:Molting / Q000235:genetics*; D010641:Phenotype; D017354:Point Mutation / Q000235:genetics; D012333:RNA, Messenger / Q000235:genetics / Q000378:metabolism*; D016415:Sequence Alignment; D017422:Sequence Analysis, DNA
D017931:DNA Primers; D007447:Invertebrate Hormones; D012333:RNA, Messenger; C002295:bursicon