Formation of Rigid, Non-Flight Forewings (Elytra) of a Beetle Requires Two Major Cuticular Proteins

English version České info

Insect cuticle is composed primarily of chitin and structural proteins. To study the function of structural cuticular proteins, we focused on the proteins present in elytra (modified forewings that become highly sclerotized and pigmented covers for the hindwings) of the red flour beetle, Tribolium castaneum. We identified two highly abundant proteins, TcCPR27 (10 kDa) and TcCPR18 (20 kDa), which are also present in pronotum and ventral abdominal cuticles. Both are members of the Rebers and Riddiford family of cuticular proteins and contain RR2 motifs. Transcripts for both genes dramatically increase in abundance at the pharate adult stage and then decline quickly thereafter. Injection of specific double-stranded RNAs for each gene into penultimate or last instar larvae had no effect on larval–larval, larval–pupal, or pupal–adult molting. The elytra of the resulting adults, however, were shorter, wrinkled, warped, fenestrated, and less rigid than those from control insects. TcCPR27-deficient insects could not fold their hindwings properly and died prematurely approximately one week after eclosion, probably because of dehydration. TcCPR18-deficient insects exhibited a similar but less dramatic phenotype. Immunolocalization studies confirmed the presence of TcCPR27 in the elytral cuticle. These results demonstrate that TcCPR27 and TcCPR18 are major structural proteins in the rigid elytral, dorsal thoracic, and ventral abdominal cuticles of the red flour beetle, and that both proteins are required for morphogenesis of the beetle's elytra.

Vyšlo v časopise: Formation of Rigid, Non-Flight Forewings (Elytra) of a Beetle Requires Two Major Cuticular Proteins. PLoS Genet 8(4): e32767. doi:10.1371/journal.pgen.1002682
Kategorie: Research Article
prolekare.web.journal.doi_sk: https://doi.org/10.1371/journal.pgen.1002682

Souhrn

Zdroje

1. MoussianB 2010 Recent advances in understanding mechanisms of insect cuticle differentiation. Insect Biochem Mol Biol 40 363 375

2. ArakaneYMuthukrishnanSBeemanRWKanostMRKramerKJ 2005 Laccase 2 is the phenoloxidase gene required for beetle cuticle tanning. Proc Natl Acad Sci U S A 102 11337 11342

3. HopkinsTLKramerKJ 1992 Insect cuticle sclerotization. Annual Review of Entomology 37 273 302

4. KarouzouMVSpyropoulosYIconomidouVACornmanRSHamodrakasSJ 2007 Drosophila cuticular proteins with the R&R Consensus: annotation and classification with a new tool for discriminating RR-1 and RR-2 sequences. Insect Biochem Mol Biol 37 754 760

5. RichardsSGibbsRAWeinstockGMBrownSJDenellR 2008 The genome of the model beetle and pest Tribolium castaneum. Nature 452 949 955

6. FutahashiROkamotoSKawasakiHZhongYSIwanagaM 2008 Genome-wide identification of cuticular protein genes in the silkworm, Bombyx mori. Insect Biochem Mol Biol 38 1138 1146

7. WillisJH 2010 Structural cuticular proteins from arthropods: annotation, nomenclature, and sequence characteristics in the genomics era. Insect Biochem Mol Biol 40 189 204

8. CornmanRSTogawaTDunnWAHeNEmmonsAC 2008 Annotation and analysis of a large cuticular protein family with the R&R Consensus in Anopheles gambiae. BMC Genomics 9 22

9. CornmanRSWillisJH 2009 Annotation and analysis of low-complexity protein families of Anopheles gambiae that are associated with cuticle. Insect Mol Biol 18 607 622

10. TogawaTAugustine DunnWEmmonsACWillisJH 2007 CPF and CPFL, two related gene families encoding cuticular proteins of Anopheles gambiae and other insects. Insect Biochem Mol Biol 37 675 688

11. RebersJERiddifordLM 1988 Structure and expression of a Manduca sexta larval cuticle gene homologous to Drosophila cuticle genes. J Mol Biol 203 411 423

12. TogawaTNakatoHIzumiS 2004 Analysis of the chitin recognition mechanism of cuticle proteins from the soft cuticle of the silkworm, Bombyx mori. Insect Biochem Mol Biol 34 1059 1067

13. RebersJEWillisJH 2001 A conserved domain in arthropod cuticular proteins binds chitin. Insect Biochem Mol Biol 31 1083 1093

14. HuntTBergstenJLevkanicovaZPapadopoulouAJohnOS 2007 A comprehensive phylogeny of beetles reveals the evolutionary origins of a superradiation. Science 318 1913 1916

15. GrimaldiDEngelMS 2005 Evolution of the Insects. Cambridge University Press ISBN 0-521-82149-5

16. Prud'hommeBMinervinoCHocineMCandeJDAouaneA 2011 Body plan innovation in treehoppers through the evolution of an extra wing-like appendage. Nature 473 83 86

17. ChenJDaiGXuYIwamotoM 2007 Optimal composite structures in the forewings of beetles. Composite Structures 81 432 437

18. ChenJNiQQXuYIwamotoM 2007 Lightweight composite structures in the forewings of beetles. Composite Structures 79 331 337

19. LomakinJHuberPAEichlerCArakaneYKramerKJ 2011 Mechanical properties of the beetle elytron, a biological composite material. Biomacromolecules 12 321 335

20. BouhinHCharlesJPQuennedeyBDelachambreJ 1992 Developmental profiles of epidermal mRNAs during the pupal-adult molt of Tenebrio molitor and isolation of a cDNA clone encoding an adult cuticular protein: effects of a juvenile hormone analogue. Dev Biol 149 112 122

21. DittmerNTHiromasaYTomichJMLuNBeemanRW 2011 Proteomic and transcriptomic analyses of rigid and membranous cuticles and epidermis from the elytra and hindwings of the red flour beetle, Tribolium castaneum. J Proteome Res In press

22. LorenzenMDBrownSJDenellREBeemanRW 2002 Cloning and characterization of the Tribolium castaneum eye-color genes encoding tryptophan oxygenase and kynurenine 3-monooxygenase. Genetics 160 225 234

23. FriedJR 2003 Polymer Science and Technology, 2nd ed Prentice Hall: Upper Saddle River, NJ

24. SperlingLH 2006 Introduction to Physical Polymer Science, 4th ed Wiley- Interscience, Hoboken, NJ

25. ErmanB 1997 Structures and properties of rubberlike network Oxford University Press: New York

26. GehrkeSH 2000 Synthesis and properties of hydrogels used for drug delivery. AmidonGLLeePIToppEM Transport Processes in Pharmaceutical Systems 473 546

27. HershBMNelsonCEStollSJNortonJEAlbertTJ 2007 The UBX-regulated network in the haltere imaginal disc of D. melanogaster. Dev Biol 302 717 727

28. TomoyasuYWheelerSRDenellRE 2005 Ultrabithorax is required for membranous wing identity in the beetle Tribolium castaneum. Nature 433 643 647

29. TomoyasuYArakaneYKramerKJDenellRE 2009 Repeated co-options of exoskeleton formation during wing-to-elytron evolution in beetles. Curr Biol 19 2057 2065

30. MissiosSDavidsonHCLinderDMortimerLOkobiAO 2000 Characterization of cuticular proteins in the red flour beetle, Tribolium castaneum. Insect Biochem Mol Biol 30 47 56

31. LampeDJWillisJH 1994 Characterization of a cDNA and gene encoding a cuticular protein from rigid cuticles of the giant silkmoth, Hyalophora cecropia. Insect Biochem Mol Biol 24 419 435

32. KramerKJKanostMRHopkinsTLJiangHZhuYC 2001 Oxidative conjugation of catechols with proteins in insect skeletal systems. Tetrahedon 57 385 392

33. MiserezARubinDWaiteJH 2010 Cross-linking chemistry of squid beak. J Biol Chem 285 38115 38124

34. LomakinJArakaneYKramerKJBeemanRWKanostMR 2010 Mechanical properties of elytra from Tribolium castaneum wild-type and body color mutant strains. J Insect Physiol 56 1901 1906

35. AndersenSO 2012 Cuticular Sclerotization and Tanning: Insect Molecular Biology and Biochemistry. GilbertLI Elsevier 167 192

36. BeemanRWStuartJJ 1990 A gene for lindane+cyclodiene resistance in the red flour beetle (Coleoptera: Tenebrionidae). J Econ Entomol 83 1745 1751

37. ArakaneYDixitRBegumKParkYSpechtCA 2009 Analysis of functions of the chitin deacetylase gene family in Tribolium castaneum. Insect Biochem Mol Biol 39 355 365

38. ArakaneYMuthukrishnanSKramerKJSpechtCATomoyasuY 2005 The Tribolium chitin synthase genes TcCHS1 and TcCHS2 are specialized for synthesis of epidermal cuticle and midgut peritrophic matrix. Insect Mol Biol 14 453 463

39. TomoyasuYDenellRE 2004 Larval RNAi in Tribolium (Coleoptera) for analyzing adult development. Dev Genes Evol 214 575 578

40. ArakaneYLomakinJBeemanRWMuthukrishnanSGehrkeSH 2009 Molecular and functional analyses of amino acid decarboxylases involved in cuticle tanning in Tribolium castaneum. J Biol Chem 284 16584 16594

41. GormanMJArakaneY 2010 Tyrosine hydroxylase is required for cuticle sclerotization and pigmentation in Tribolium castaneum. Insect Biochem Mol Biol 40 267 273