Laccase2 is required for cuticular pigmentation in stinkbugs

Futahashi, Ryo; Tanaka, Kohjiro; Matsuura, Yu; Tanahashi, Masahiko; Kikuchi, Yoshitomo; Fukatsu, Takema

doi:10.1016/j.ibmb.2010.12.003

Cited by 87 publications

(72 citation statements)

References 33 publications

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“…1A, right). Notably, unlike the PlexA locus, the 490-nt Laccase2 transcript was more abundant (approximately fivefold and ∼2.5-fold in DL1 and S2 cells, respectively) than the linear Laccase2 mRNA, which encodes an enzyme implicated in cuticle formation and pigmentation (Futahashi et al 2011;Jacobs et al 2015). Using RT-PCR (Supplemental Fig.…”

Section: Resultsmentioning

confidence: 99%

Combinatorial control of Drosophila circular RNA expression by intronic repeats, hnRNPs, and SR proteins

et al. 2015

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Thousands of eukaryotic protein-coding genes are noncanonically spliced to produce circular RNAs. Bioinformatics has indicated that long introns generally flank exons that circularize in Drosophila, but the underlying mechanisms by which these circular RNAs are generated are largely unknown. Here, using extensive mutagenesis of expression plasmids and RNAi screening, we reveal that circularization of the Drosophila laccase2 gene is regulated by both intronic repeats and trans-acting splicing factors. Analogous to what has been observed in humans and mice, basepairing between highly complementary transposable elements facilitates backsplicing. Long flanking repeats (∼400 nucleotides [nt]) promote circularization cotranscriptionally, whereas pre-mRNAs containing minimal repeats (<40 nt) generate circular RNAs predominately after 3 ′ end processing. Unlike the previously characterized Muscleblind (Mbl) circular RNA, which requires the Mbl protein for its biogenesis, we found that Laccase2 circular RNA levels are not controlled by Mbl or the Laccase2 gene product but rather by multiple hnRNP (heterogeneous nuclear ribonucleoprotein) and SR (serine-arginine) proteins acting in a combinatorial manner. hnRNP and SR proteins also regulate the expression of other Drosophila circular RNAs, including Plexin A (PlexA), suggesting a common strategy for regulating backsplicing. Furthermore, the laccase2 flanking introns support efficient circularization of diverse exons in Drosophila and human cells, providing a new tool for exploring the functional consequences of circular RNA expression across eukaryotes.

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Section: Resultsmentioning

confidence: 99%

Combinatorial control of Drosophila circular RNA expression by intronic repeats, hnRNPs, and SR proteins

et al. 2015

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“…and Kleidocerys spp. ; (2) the Lygaeidae also embraces stinkbugs without the bacteriome such as Spilostethus, Graptostethus and Tropidothorax species; (3) therefore, morphological, developmental and molecular biological comparisons between allied species with and without the symbiotic organ are feasible; (4) in Nysius plebeius and Oncopeltus fasciatus, it has been shown that RNA interference works efficiently (Hughes and Kaufman, 2000;Futahashi et al, 2011), which enables molecular genetic approaches to the mechanisms as to what host genes are involved in the endosymbiosis and how the symbiotic organ is differentiated and formed; and (5) we have established rearing techniques for the Nysius, Kleidocerys, Spilostethus and Graptostethus species in Petri dishes on plant seeds, which offer tractable model systems for experimental and functional studies. Thus far, the aphid-Buchnera association has been the best-studied model endosymbiotic system with the bacteriome, wherein both the host genome and the endosymbiont genome have been sequenced (Shigenobu et al, 2000;International Aphid Genomics Consortium, 2010a), developmental processes of the bacteriome formation have been histologically documented Miura et al, 2003), the transcriptomics of the symbiotic organ have been conducted (Nakabachi et al, 2005;Hansen and Moran, 2011) and a number of physiological, ecological and molecular works on the endosymbiosis have been accumulated (reviewed in Douglas, 1998;Baumann, 2005).…”

Section: Perspectivesmentioning

confidence: 99%

Evolution of symbiotic organs and endosymbionts in lygaeid stinkbugs

et al. 2011

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We investigated seed bugs of the genus Nysius (Insecta: Hemiptera: Lygaeidae) for their symbiotic bacteria. From all the samples representing 4 species, 18 populations and 281 individuals, specific bacterial 16S rRNA gene sequences were consistently identified, which formed a distinct clade in the Gammaproteobacteria. In situ hybridization showed that the bacterium was endocellularly localized in a pair of large bacteriomes that were amorphous in shape, deep red in color, and in association with gonads. In the ovary of adult females, the endosymbiont was also localized in the 'infection zone' in the middle of each germarium and in the 'symbiont ball' at the anterior pole of each oocyte, indicating vertical transmission of the endosymbiont through the ovarial passage. Phylogenetic analyses based on bacterial 16S rRNA, groEL and gyrB genes consistently supported a coherent monophyly of the Nysius endosymbionts. The possibility of a sister relationship to 'Candidatus Kleidoceria schneideri', the bacteriome-associated endosymbiont of a lygaeid bug Kleidocerys resedae, was statistically rejected, indicating independent evolutionary origins of the endosymbionts in the Lygaeidae. The endosymbiont genes consistently exhibited AT-biased nucleotide compositions and accelerated rates of molecular evolution, and the endosymbiont genome was only 0.6 Mb in size. The endosymbiont phylogeny was congruent with the host insect phylogeny, suggesting strict vertical transmission and host-symbiont co-speciation over evolutionary time. Based on these results, we discuss the evolution of bacteriomes and endosymbionts in the Heteroptera, most members of which are associated with gut symbiotic bacteria. The designation 'Candidatus Schneideria nysicola' is proposed for the endosymbiont clade.

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“…An initial demonstration of RNAi in pentatomid stink bugs, by injection, would support this insect's capacity to elicit an RNAi response and confirm that stink bugs possess the cellular machinery necessary for RNAi. Injection-based RNAi responses have been described in other families of Heteroptera; Futahashi et al (2011) describe the role of Laccase2 in cuticular pigment production of bean bug, Riptortus pedestris (Hemiptera: Alydidae), seed bug, Nysius plebeius (Hemiptera: Lygaeidae), and a Kudzu Bug relative, Japanese common plataspid stinkbug, Megacopta punctatissima (Hemiptera: Plataspididae).…”

Section: Introductionmentioning

confidence: 99%