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      Analysis and Functional Annotation of Expressed Sequence Tags from the Asian Longhorned Beetle, Anoplophora glabripennis

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          Abstract

          The Asian longhorned beetle, Anoplophora glabripennis (Motschulsky) (Coleoptera: Cerambycidae), is one of the most economically and ecologically devastating forest insects to invade North America in recent years. Despite its substantial impact, limited effort has been expended to define the genetic and molecular make-up of this species. Considering the significant role played by late-stadia larvae in host tree decimation, a small-scale EST sequencing project was done using a cDNA library constructed from 5 th -instar A. glabripennis. The resultant dataset consisted of 599 high quality ESTs that, upon assembly, yielded 381 potentially unique transcripts. Each of these transcripts was catalogued as to putative molecular function, biological process, and associated cellular component according to the Gene Ontology classification system. Using this annotated dataset, a subset of assembled sequences was identified that are putatively associated with A. glabnpennis development and metamorphosis. This work will contribute to understanding of the diverse molecular mechanisms that underlie coleopteran morphogenesis and enable the future development of novel control strategies for management of this insect pest.

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          Reconstitution of actin-based motility of Listeria and Shigella using pure proteins.

          Actin polymerization is essential for cell locomotion and is thought to generate the force responsible for cellular protrusions. The Arp2/3 complex is required to stimulate actin assembly at the leading edge in response to signalling. The bacteria Listeria and Shigella bypass the signalling pathway and harness the Arp2/3 complex to induce actin assembly and to propel themselves in living cells. However, the Arp2/3 complex alone is insufficient to promote movement. Here we have used pure components of the actin cytoskeleton to reconstitute sustained movement in Listeria and Shigella in vitro. Actin-based propulsion is driven by the free energy released by ATP hydrolysis linked to actin polymerization, and does not require myosin. In addition to actin and activated Arp2/3 complex, actin depolymerizing factor (ADF, or cofilin) and capping protein are also required for motility as they maintain a high steady-state level of G-actin, which controls the rate of unidirectional growth of actin filaments at the surface of the bacterium. The movement is more effective when profilin, alpha-actinin and VASP (for Listeria) are also included. These results have implications for our understanding of the mechanism of actin-based motility in cells.
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            Exotic bark- and wood-boring Coleoptera in the United States: recent establishments and interceptions

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              Identification of Drosophila melanogaster yellow-f and yellow-f2 proteins as dopachrome-conversion enzymes.

              This study describes the identification of Drosophila yellow-f and yellow-f2 as dopachrome-conversion enzymes responsible for catalysing the conversion of dopachrome into 5,6-dihydroxyindole in the melanization pathway. Drosophila yellow -y gene and yellow -b, -c, -f and -f2 genes were expressed in an insect cell/baculovirus expression system and their corresponding recombinant proteins were screened for dopachrome-conversion enzyme activity. Among the yellow and yellow -related genes, the yellow -f and yellow -f2 genes were identified as the genes coding for Drosophila dopachrome-conversion enzyme based on the high activity of their recombinant proteins in catalysing the production of 5,6-dihydroxyindole from dopachrome. Both yellow-f and yellow-f2 are capable of mediating a decarboxylative structural rearrangement of dopachrome, as well as an isomerization/tautomerization of dopamine chrome and dopa methyl ester chrome. Northern hybridization revealed the transcription of yellow -f in larvae and pupae, but a high abundance of mRNA was observed in later larval and early pupal stages. In contrast, yellow-f2 transcripts were present at all stages, but high abundance of its mRNA was observed in later-stage pupae and adults. These data indicate that yellow-f and yellow-f2 complement each other during Drosophila development and that the yellow-f is involved in larval and pupal melanization, and yellow-f2 plays a major role in melanization reactions in Drosophila during later pupal and adult development. Results from this study provide the groundwork towards a better understanding of the physiological roles of the Drosophila yellow gene family.
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                Author and article information

                Journal
                J Insect Sci
                J. Insect Sci
                insc
                Journal of Insect Science
                University of Wisconsin Library
                1536-2442
                2009
                20 May 2009
                : 9
                : 21
                Affiliations
                [ 1 ]U.S. Department of Agriculture, Agricultural Research Service, U.S. Horticultural Research Laboratory, Ft. Pierce, Florida 34945
                [ 2 ]U.S. Department of Agriculture, Agricultural Research Service, Beneficial Insects Introduction Research Unit, Newark, Delaware 19713
                [ 3 ]Current address: North Carolina State University, Raleigh, North Carolina 27695
                Author notes
                Article
                10.1673/031.009.2101
                3011843
                19619025
                788fdeba-7ffe-47af-850b-ddd56e3bcb31
                © 2009

                This is an open-access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited.

                History
                : 3 September 2007
                : 8 March 2008
                Page count
                Pages: 13
                Categories
                Article

                Entomology
                morphogenesis,development,insect,coleoptera,transcriptome,est
                Entomology
                morphogenesis, development, insect, coleoptera, transcriptome, est

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