Cloning and expression analysis of a G‐protein subunit— in the rice water weevil lissorhoptrus oryzophilus Kuschel

Abstract

The open reading frame (ORF) encoding a novel G protein α subunit, Lo Gα(o), was cloned from the parthenogenetic rice water weevil, Lissorhoptrus oryzophilus Kuschel (Coleoptera: Curculionidae). The Lo Gα(o) ORF encodes a protein of 354 amino acid residues. The deduced protein sequence shares high homology with Gα(o) from other species. The expression patterns of Lo Gα(o) in various adult tissues were indicated by real-time quantitative PCR and Western blot. The results showed that Lo Gα(o) mRNA was expressed at similar levels in tissues except relative high levels in the antennae of adult, and Lo Gα(o) protein of an apparent molecular mass of about 40 kDa was expressed in various tissues of the adult. Immunocytochemical localization showed that Lo Gα(o) was mainly expressed in the dendrites of the trichoid sensilla in the antenna of the weevil. The tissue and cellular localization of Lo Gα(o) suggests that Lo Gα(o) may take a part in signal transduction of olfactory/gustatory.

Role of Go/i subgroup of G proteins in olfactory signaling of Drosophila melanogaster

“…Insect ORNs express several G proteins that could be involved in signal transduction, in particular the G o/i subgroup of G proteins (Miura et al ., 2005 ; Rutzler et al ., 2006 ; Boto et al ., 2010 ; Kang et al ., 2011 ). Therefore, in this study we tested whether G o/i are required for olfaction in behavior, for odor responses in the native tissue (antenna; in vivo ) or when expressed in a heterologous cell-culture system (HEK293T cells: Human Embryonic Kidney 293T cells; in vitro ).…”

CLONING AND EXPRESSION ANALYSIS OF G‐PROTEIN Gαq SUBUNIT AND Gβ1 SUBUNIT FROM Bemisia tabaci GENNADIUS (HOMOPTERA: ALEYRODIDAE)

“…In the Drosophila melanogaster genome, six genes have been described to encode the Gα proteins: Gs, Gi, Gq, Go, Gf, and concertina (cta) (Boto et al., ). In addition, there are several G protein α subunits that had been cloned from other insect species including Gαq from Anopheles gambiae (Rutzler et al., ), Bombyx mori (Miura et al., ), Helicoverpa armigera (Su et al., ), Helicoverpa assulta (Qiao et al., ), Sitobion avena (Fan et al., ), Mamestra brassicae (Jacquin‐Joly et al., ); Gαs from A. gambiae (Rutzler et al., ), B. mori (Miura et al., ; Hull et al., ); Gαi from A. gambiae (Rutzler et al., ), Manduca sexta (Horgan et al., ); Gαo from A. gambiae (Rutzler et al., ), B. mori (Miura et al., ), M. sexta (Horgan et al., ), Locusta migratoria (Raming et al., ), Lissorhoptrus oryzophilus (Kang et al., ); Gα 73B from B. mori (Zhang et al., ); Gα12 from B. mori (Zhang et al., ). The G protein β and γ subunits form a functional unit that is not dissociable except by denaturation (Schmidt et al., ; Offermanns, ).…”

Molecular Cloning and Characterization of G Alpha Proteins from the Western Tarnished Plant Bug, Lygus hesperus

“…A majority of the LhGα transcripts were amplified from all of the tissue sets from both sexes, albeit to varying degrees. Similarly wide tissue distribution profiles for Gα subunits have been reported in B. mori [ 19 ], A. gambiae [ 20 ], D. melanogaster [ 21 ], B. tabaci [ 57 ], and L. oryzophilus [ 55 ] and likely reflect the critical role of G proteins in mediating the diverse signal transduction cascades that drive cellular processes. LhGαq4 was the lone LhGα transcript to exhibit tissue specific expression with amplification limited to head-derived cDNAs ( Figure 5 B).…”

“…The expression of Gαs in olfactory neurons coupled with abnormal olfactory behavior following disruption of the Gαs signal transduction cascade [ 25 ] has led some to postulate that Gαs functions in olfaction. However, elevated levels of Gαo and Gαq transcripts have been reported in antennae and olfactory neurons of a number of insects [ 17 , 18 , 19 , 54 , 55 ]. Furthermore, similar to the Gαs pathway, downstream effectors of Gαq such as Ca 2+ /calmodulin can also activate adenylyl cyclase [ 79 ] and RNAi-mediated knockdown of Gαq likewise reduces antennal responses [ 29 ].…”

“…complex, transcriptional resources have only recently been developed [ 43 , 44 , 45 , 46 ], and our knowledge of chemosensory signal transduction is limited to odorant binding proteins [ 45 , 47 ] and the olfactory receptor co-receptor (Orco) [ 12 ] in L. lineolaris and L. hesperus . Furthermore, while G proteins have been studied in a number of insects with Gα subunits cloned from Drosophila melanogaster [ 17 , 21 , 48 , 49 , 50 ], Anopheles gambiae [ 20 ], Bombyx mori [ 19 , 51 , 52 ], Manduca sexta [ 53 ], Locusta migratoria [ 54 ], Lissorhoptrus oryzophilus [ 55 ], Helicoverpa assaulta [ 56 ], Mamestra brassicae [ 18 ], Bemisia tabaci [ 57 ], and Oncopeltus fasciatus [ 58 ], little progress has been made on the role of these genes in mediating chemosensory behaviors in plant bugs such as Lygus . In this study, we sought to begin to address this lack of knowledge by identifying the molecular sequences and expression profile of Gα subunits in L. hesperus .…”