Molecular and functional characterisation of resilin across three insect orders

Lyons, Russell E., Wong, Darren C. C., Kim, Misook, Lekieffre, Nicolas, Huson, Mickey G., Vuocolo, Tony, Merritt, David J., Nairn, Kate M., Dudek, Daniel M., Colgrave, Michelle L. and Elvin, Christopher M. (2011) Molecular and functional characterisation of resilin across three insect orders. Insect Biochemistry and Molecular Biology, 41 11: 881-890. doi:10.1016/j.ibmb.2011.08.002


Author Lyons, Russell E.
Wong, Darren C. C.
Kim, Misook
Lekieffre, Nicolas
Huson, Mickey G.
Vuocolo, Tony
Merritt, David J.
Nairn, Kate M.
Dudek, Daniel M.
Colgrave, Michelle L.
Elvin, Christopher M.
Title Molecular and functional characterisation of resilin across three insect orders
Journal name Insect Biochemistry and Molecular Biology   Check publisher's open access policy
ISSN 0965-1748
1879-0240
Publication date 2011-11-01
Year available 2011
Sub-type Article (original research)
DOI 10.1016/j.ibmb.2011.08.002
Open Access Status Not yet assessed
Volume 41
Issue 11
Start page 881
End page 890
Total pages 10
Place of publication Oxford, United Kingdom
Publisher Pergamon
Language eng
Subject 1303 Biochemistry
1312 Molecular Biology
1109 Insect Science
Abstract Resilin is an important elastomeric protein of insects, with roles in the storage and release of energy during a variety of different functional categories including flight and jumping. To date, resilin genes and protein function have been characterised only in a small number of flying insects, despite their importance in fleas and other jumping insects. Microscopy and immunostaining studies of resilin in flea demonstrate the presence of resilin pads in the pleural arch at the top of the hind legs, a region responsible for the flea's jumping ability. A degenerate primer approach was used to amplify resilin gene transcripts from total RNA isolated from flea (Ctenocephalides fells). buffalo fly (Haematobia irritans exigua) and dragonfly (Aeshna sp.) pharate adults, and full-length transcripts were successfully isolated. Two isoforms (A and B) were amplified from each of flea and buffalo fly, and isoform B only in dragonfly. Flea and buffalo fly isoform B transcripts were expressed in an Escherichia coli expression system, yielding soluble recombinant proteins Cf-resB and Hi-resB respectively. Protein structure and mechanical properties of each protein before and after crosslinking were assessed. This study shows that resilin gene and protein sequences are broadly conserved and that crosslinked recombinant resilin proteins share similar mechanical properties from flying to jumping insects. A combined use of degenerate primers and polyclonal sera will likely facilitate characterisation of resilin genes from other insect and invertebrate orders. Crown Copyright (C) 2011 Published by Elsevier Ltd. All rights reserved.
Formatted abstract
Resilin is an important elastomeric protein of insects, with roles in the storage and release of energy during a variety of different functional categories including flight and jumping. To date, resilin genes and protein function have been characterised only in a small number of flying insects, despite their importance in fleas and other jumping insects. Microscopy and immunostaining studies of resilin in flea demonstrate the presence of resilin pads in the pleural arch at the top of the hind legs, a region responsible for the flea’s jumping ability. A degenerate primer approach was used to amplify resilin gene transcripts from total RNA isolated from flea (Ctenocephalides felis), buffalo fly (Haematobia irritans exigua) and dragonfly (Aeshna sp.) pharate adults, and full-length transcripts were successfully isolated. Two isoforms (A and B) were amplified from each of flea and buffalo fly, and isoform B only in dragonfly. Flea and buffalo fly isoform B transcripts were expressed in an Escherichia coli expression system, yielding soluble recombinant proteins Cf-resB and Hi-resB respectively. Protein structure and mechanical properties of each protein before and after crosslinking were assessed. This study shows that resilin gene and protein sequences are broadly conserved and that crosslinked recombinant resilin proteins share similar mechanical properties from flying to jumping insects. A combined use of degenerate primers and polyclonal sera will likely facilitate characterisation of resilin genes from other insect and invertebrate orders.
Keyword Biochemistry & Molecular Biology
Entomology
Biochemistry & Molecular Biology
Entomology
BIOCHEMISTRY & MOLECULAR BIOLOGY
ENTOMOLOGY
Q-Index Code C1
Q-Index Status Confirmed Code
Institutional Status UQ

Document type: Journal Article
Sub-type: Article (original research)
Collections: Official 2012 Collection
School of Biological Sciences Publications
 
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Citation counts: TR Web of Science Citation Count  Cited 24 times in Thomson Reuters Web of Science Article | Citations
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Created: Tue, 18 Oct 2011, 21:32:34 EST by Gail Walter on behalf of School of Biological Sciences