Skip to main content
PMC home page
Biochemical Journal logoLink to Biochemical Journal
. 2002 Oct 1;367(Pt 1):187–193. doi: 10.1042/BJ20020567

Ance, a Drosophila angiotensin-converting enzyme homologue, is expressed in imaginal cells during metamorphosis and is regulated by the steroid, 20-hydroxyecdysone.

Richard J Siviter 1, Christine A M Taylor 1, Deborah M Cottam 1, Adrian Denton 1, M Paulina Dani 1, Martin J Milner 1, Alan D Shirras 1, R Elwyn Isaac 1
PMCID: PMC1222869  PMID: 12093364

Abstract

Ance is a single domain homologue of mammalian angiotensin-converting enzyme (ACE) and is important for normal development and reproduction in Drosophila melanogaster. Mammalian ACE is responsible for the synthesis of angiotensin II and the inactivation of bradykinin and N -acetyl-Ser-Asp-Lys-Pro, but the absence of similar peptide hormones in insects suggests novel functions for Ance. We now provide evidence in support of a role for Ance during Drosophila metamorphosis. The transition of larva to pupa was accompanied by a 3-fold increase in ACE-like activity, which subsequently dropped to larval levels on adult eclosion. This increase was attributed to the induction of Ance expression during the wandering phase of the last larval instar in the imaginal cells (imaginal discs, abdominal histoblasts, gut imaginal cells and imaginal salivary gland). Ance expression was particularly strong in the presumptive adult midgut formed as a result of massive proliferation of the imaginal midgut cells soon after pupariation. No Ance transcripts were detected in the midgut of the fully differentiated adult intestine. Ance protein and mRNA were not detected in imaginal discs from wandering larvae of flies homozygous for the ecd ( 1 ) allele, a temperature-sensitive ecdysone-less mutant, suggesting that Ance expression is ecdysteroid-dependent. Physiological levels of 20-hydroxyecdysone induced the synthesis of ACE-like activity and Ance protein by a wing disc cell line (Cl.8+), confirming that Ance is an ecdysteroid-responsive gene. We propose that the expression of Ance in imaginal cells is co-ordinated by exposure to ecdysteroid (moulting hormone) during the last larval instar moult to increase levels of ACE-like activity during metamorphosis. The enzyme activity may be required for the processing of a developmental peptide hormone or may function in concert with other peptidases to provide amino acids for the synthesis of adult proteins.

Full Text

The Full Text of this article is available as a PDF (227.6 KB).

Selected References

These references are in PubMed. This may not be the complete list of references from this article.

  1. Amin A., Li Y., Finkelstein R. Identification of a Drosophila prolyl endopeptidase and analysis of its expression. DNA Cell Biol. 1999 Aug;18(8):605–610. doi: 10.1089/104454999315015. [DOI] [PubMed] [Google Scholar]
  2. Andres A. J., Thummel C. S. Methods for quantitative analysis of transcription in larvae and prepupae. Methods Cell Biol. 1994;44:565–573. doi: 10.1016/s0091-679x(08)60932-2. [DOI] [PubMed] [Google Scholar]
  3. Appel L. F., Prout M., Abu-Shumays R., Hammonds A., Garbe J. C., Fristrom D., Fristrom J. The Drosophila Stubble-stubbloid gene encodes an apparent transmembrane serine protease required for epithelial morphogenesis. Proc Natl Acad Sci U S A. 1993 Jun 1;90(11):4937–4941. doi: 10.1073/pnas.90.11.4937. [DOI] [PMC free article] [PubMed] [Google Scholar]
  4. Ashe H. L., Levine M. Local inhibition and long-range enhancement of Dpp signal transduction by Sog. Nature. 1999 Apr 1;398(6726):427–431. doi: 10.1038/18892. [DOI] [PubMed] [Google Scholar]
  5. Berreur P., Porcheron P., Moriniere M., Berreur-Bonnenfant J., Belinski-Deutsch S., Busson D., Lamour-Audit C. Ecdysteroids during the third larval instar in 1(3)ecd-1ts, a temperature-sensitive mutant of Drosophila melanogaster. Gen Comp Endocrinol. 1984 Apr;54(1):76–84. doi: 10.1016/0016-6480(84)90201-6. [DOI] [PubMed] [Google Scholar]
  6. Cornell M. J., Williams T. A., Lamango N. S., Coates D., Corvol P., Soubrier F., Hoheisel J., Lehrach H., Isaac R. E. Cloning and expression of an evolutionary conserved single-domain angiotensin converting enzyme from Drosophila melanogaster. J Biol Chem. 1995 Jun 9;270(23):13613–13619. doi: 10.1074/jbc.270.23.13613. [DOI] [PubMed] [Google Scholar]
  7. Corvol P., Williams T. A., Soubrier F. Peptidyl dipeptidase A: angiotensin I-converting enzyme. Methods Enzymol. 1995;248:283–305. doi: 10.1016/0076-6879(95)48020-x. [DOI] [PubMed] [Google Scholar]
  8. Hall N. A. Peptidases in Drosophila melanogaster. I. Characterization of dipeptidase and leucine aminopeptidase activities. Biochem Genet. 1986 Oct;24(9-10):775–793. doi: 10.1007/BF00499009. [DOI] [PubMed] [Google Scholar]
  9. Haunerland N. H. Insect storage proteins: gene families and receptors. Insect Biochem Mol Biol. 1996 Sep-Oct;26(8-9):755–765. doi: 10.1016/s0965-1748(96)00035-5. [DOI] [PubMed] [Google Scholar]
  10. Henrich V. C., Tucker R. L., Maroni G., Gilbert L. I. The ecdysoneless (ecd1ts) mutation disrupts ecdysteroid synthesis autonomously in the ring gland of Drosophila melanogaster. Dev Biol. 1987 Mar;120(1):50–55. doi: 10.1016/0012-1606(87)90102-3. [DOI] [PubMed] [Google Scholar]
  11. Houard X., Williams T. A., Michaud A., Dani P., Isaac R. E., Shirras A. D., Coates D., Corvol P. The Drosophila melanogaster-related angiotensin-I-converting enzymes Acer and Ance--distinct enzymic characteristics and alternative expression during pupal development. Eur J Biochem. 1998 Nov 1;257(3):599–606. doi: 10.1046/j.1432-1327.1998.2570599.x. [DOI] [PubMed] [Google Scholar]
  12. Isaac R. E., Ekbote U., Coates D., Shirras A. D. Insect angiotensin-converting enzyme. A processing enzyme with broad substrate specificity and a role in reproduction. Ann N Y Acad Sci. 1999;897:342–347. doi: 10.1111/j.1749-6632.1999.tb07904.x. [DOI] [PubMed] [Google Scholar]
  13. Isaac R. E., Lamango N. S. Peptidyl dipeptidase activity in the haemolymph of insects. Biochem Soc Trans. 1994 Aug;22(3):292S–292S. doi: 10.1042/bst022292s. [DOI] [PubMed] [Google Scholar]
  14. Isaac R., Schoofs L., Williams T. A., Veelaert D., Sajid M., Corvol P., Coates D. A novel peptide-processing activity of insect peptidyl-dipeptidase A (angiotensin I-converting enzyme): the hydrolysis of lysyl-arginine and arginyl-arginine from the C-terminus of an insect prohormone peptide. Biochem J. 1998 Feb 15;330(Pt 1):61–65. doi: 10.1042/bj3300061. [DOI] [PMC free article] [PubMed] [Google Scholar]
  15. Jiang C., Baehrecke E. H., Thummel C. S. Steroid regulated programmed cell death during Drosophila metamorphosis. Development. 1997 Nov;124(22):4673–4683. doi: 10.1242/dev.124.22.4673. [DOI] [PubMed] [Google Scholar]
  16. Laemmli U. K. Cleavage of structural proteins during the assembly of the head of bacteriophage T4. Nature. 1970 Aug 15;227(5259):680–685. doi: 10.1038/227680a0. [DOI] [PubMed] [Google Scholar]
  17. Lamango N. S., Isaac R. E. Identification and properties of a peptidyl dipeptidase in the housefly, Musca domestica, that resembles mammalian angiotensin-converting enzyme. Biochem J. 1994 May 1;299(Pt 3):651–657. doi: 10.1042/bj2990651. [DOI] [PMC free article] [PubMed] [Google Scholar]
  18. Lamango N. S., Nachman R. J., Hayes T. K., Strey A., Isaac R. E. Hydrolysis of insect neuropeptides by an angiotensin-converting enzyme from the housefly, Musca domestica. Peptides. 1997;18(1):47–52. doi: 10.1016/s0196-9781(96)00232-x. [DOI] [PubMed] [Google Scholar]
  19. Lamango N., Isaac R. E. Identification of an ACE-like peptidyl dipeptidase activity in the housefly, Musca domestica. Biochem Soc Trans. 1993 Aug;21(3):245S–245S. doi: 10.1042/bst021245s. [DOI] [PubMed] [Google Scholar]
  20. Ohtsuki S., Homma K., Kurata S., Komano H., Natori S. A prolyl endopeptidase of Sarcophaga peregrina (flesh fly): its purification and suggestion for its participation in the differentiation of the imaginal discs. J Biochem. 1994 Mar;115(3):449–453. doi: 10.1093/oxfordjournals.jbchem.a124358. [DOI] [PubMed] [Google Scholar]
  21. Ohtsuki S., Homma K., Kurata S., Natori S. Molecular cloning of cDNA for Sarcophaga prolyl endopeptidase and characterization of the recombinant enzyme produced by an E. coli expression system. Insect Biochem Mol Biol. 1997 Apr;27(4):337–343. doi: 10.1016/s0965-1748(97)00004-0. [DOI] [PubMed] [Google Scholar]
  22. Pino-Heiss S., Schubiger G. Extracellular protease production by Drosophila imaginal discs. Dev Biol. 1989 Apr;132(2):282–291. doi: 10.1016/0012-1606(89)90225-x. [DOI] [PubMed] [Google Scholar]
  23. Quan G. X., Mita K., Okano K., Shimada T., Ugajin N., Xia Z., Goto N., Kanke E., Kawasaki H. Isolation and expression of the ecdysteroid-inducible angiotensin-converting enzyme-related gene in wing discs of Bombyx mori. Insect Biochem Mol Biol. 2001 Jan;31(1):97–103. doi: 10.1016/s0965-1748(00)00112-0. [DOI] [PubMed] [Google Scholar]
  24. Richards G. The radioimmune assay of ecdysteroid titres in Drosophila melanogaster. Mol Cell Endocrinol. 1981 Mar;21(3):181–197. doi: 10.1016/0303-7207(81)90013-7. [DOI] [PubMed] [Google Scholar]
  25. Rusch J., Levine M. Regulation of a dpp target gene in the Drosophila embryo. Development. 1997 Jan;124(2):303–311. doi: 10.1242/dev.124.2.303. [DOI] [PubMed] [Google Scholar]
  26. Siviter R. J., Coast G. M., Winther A. M., Nachman R. J., Taylor C. A., Shirras A. D., Coates D., Isaac R. E., Nässel D. R. Expression and functional characterization of a Drosophila neuropeptide precursor with homology to mammalian preprotachykinin A. J Biol Chem. 2000 Jul 28;275(30):23273–23280. doi: 10.1074/jbc.M002875200. [DOI] [PubMed] [Google Scholar]
  27. Tatei K., Cai H., Ip Y. T., Levine M. Race: a Drosophila homologue of the angiotensin converting enzyme. Mech Dev. 1995 Jun;51(2-3):157–168. doi: 10.1016/0925-4773(95)00349-5. [DOI] [PubMed] [Google Scholar]
  28. Taylor C. A., Coates D., Shirras A. D. The Acer gene of Drosophila codes for an angiotensin-converting enzyme homologue. Gene. 1996 Nov 28;181(1-2):191–197. doi: 10.1016/s0378-1119(96)00503-3. [DOI] [PubMed] [Google Scholar]
  29. Thummel C. S. Flies on steroids--Drosophila metamorphosis and the mechanisms of steroid hormone action. Trends Genet. 1996 Aug;12(8):306–310. doi: 10.1016/0168-9525(96)10032-9. [DOI] [PubMed] [Google Scholar]
  30. Thummel C. S. Steroid-triggered death by autophagy. Bioessays. 2001 Aug;23(8):677–682. doi: 10.1002/bies.1096. [DOI] [PubMed] [Google Scholar]
  31. Turner A. J., Isaac R. E., Coates D. The neprilysin (NEP) family of zinc metalloendopeptidases: genomics and function. Bioessays. 2001 Mar;23(3):261–269. doi: 10.1002/1521-1878(200103)23:3<261::AID-BIES1036>3.0.CO;2-K. [DOI] [PubMed] [Google Scholar]
  32. Wijffels G., Fitzgerald C., Gough J., Riding G., Elvin C., Kemp D., Willadsen P. Cloning and characterisation of angiotensin-converting enzyme from the dipteran species, Haematobia irritans exigua, and its expression in the maturing male reproductive system. Eur J Biochem. 1996 Apr 15;237(2):414–423. doi: 10.1111/j.1432-1033.1996.0414k.x. [DOI] [PubMed] [Google Scholar]
  33. Williams T. A., Michaud A., Houard X., Chauvet M. T., Soubrier F., Corvol P. Drosophila melanogaster angiotensin I-converting enzyme expressed in Pichia pastoris resembles the C domain of the mammalian homologue and does not require glycosylation for secretion and enzymic activity. Biochem J. 1996 Aug 15;318(Pt 1):125–131. doi: 10.1042/bj3180125. [DOI] [PMC free article] [PubMed] [Google Scholar]
  34. Zhao X., Mita K., Shimada T., Okano K., Quan G. X., Kanke E., Kawasaki H. Isolation and expression of an ecdysteroid-inducible neutral endopeptidase 24.11-like gene in wing discs of Bombyx mori. Insect Biochem Mol Biol. 2001 Nov 1;31(12):1213–1219. doi: 10.1016/s0965-1748(01)00069-8. [DOI] [PubMed] [Google Scholar]

Articles from Biochemical Journal are provided here courtesy of The Biochemical Society

RESOURCES