Structure and expression of mRNA for vitellogenin in Bombyx mori

doi:10.1016/0167-4781(94)90094-9

Biochimica et Biophysica Acta (BBA) - Gene Structure and Expression

Volume 1218, Issue 1, 17 May 1994, Pages 1-10

https://doi.org/10.1016/0167-4781(94)90094-9 Get rights and content

Abstract

Vitellogenin, a precursor of major yolk protein of the silkworm, Bombyx mori is a tetramer composed of each two molecules of heavy and light subunits. We cloned mRNA sequence for the B. mori vitellogenin and analyzed its structure. Sequence alignment of several overlapping cDNA clones indicated that the vitellogenin mRNA is approx. 5.7 kb, containing an open reading frame for a peptide with 1782 amino acid residues. By comparing the deduced amino acid sequence with the amino-terminal primary structures of vitellogenin subunits, it is suggested that the heavy and light subunits of the B. mori vitellogenin are encoded by a single contiguous mRNA. The primary translation product of the vitellogenin mRNA was detected in the microsomal fraction prepared from the fat body of vitellogenic females. Northern blot analysis of the fat body RNA demonstrated that the biosynthesis of vitellogenin in B. mori is regulated in a tissue-, sex- and stage-specific manner at the level of mRNA. Possible cause for discrepancy between the present results and our previous proposal (Izumi, S. and Tomino, S. (1983) Insect Biochem. 13, 81–85) on the biosynthesis of B. mori vitellogenin is also discussed.

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The in-vitro and in-vivo radio labeling unfolds the vitellogenin protein expression site of Bombyx mori L as the ventral perivisceral fat body
2023, Journal of Asia-Pacific Entomology
The present study aims to focus on the site of fat bodies involved in the differential expression of vitellogenin protein in Bombyx mori L which is not yet fully understood. Culturing fat bodies and analyzing in in vitro and in vivo approaches made it useful to study this pattern. In Bombyx mori, L. The vitellogenin protein is 200–205 kDa and resolves at two molecular weight regions that are at 180 kDa and 42 kDa. We subject dorsal perivisceral (DPV) and ventral perivisceral fat body (VPV), haemolymph for this radio labeling cell culture and incorporation experiment. Based on our in vitro and in vivo experiments of S³⁵ methionine fluorography, Scanning densitometric, Western blot analysis and RT-PCR expression studies had strongly proven that the ventral perivisceral fat body is the significant and exclusive site of differential vitellogenin expression in silkworm, Bombyx mori but not in dorsal peripheral fat body tissues, haemolymph by radiolabeling studies. The present paper describes an experimental set-up based on the incubation of ventral perivisceral and dorsal perivisceral fat bodies and the measurement of vitellogenin with S³⁵ protein labeling incorporation method. These findings from our lab were the first report proved by in vitro and in vivo labeling experiments.
Vitellogenin regulates estrogen-related receptor expression by crosstalk with the JH and IIS-TOR signaling pathway in Polyrhachis vicina Roger (Hymenoptera, Formicidae)
2021, General and Comparative Endocrinology
Citation Excerpt :
The Vg gene plays an important role in embryonic development and is the main source of nutrition for the growth and development of oocytes (Tufail and Takeda, 2008). In P. vicina; PvVg mRNA is first expressed in the egg stage, whereas in Bombyx mori and Lymantria dispar, Vg mRNA is first expressed at the larval stage (Yano et al., 1994a, 1994b; Hiremath and Lehtoma, 1997). The level of PvVg mRNA gradually changes as the ovary develops.
The Estrogen-related receptor (ERR) can regulate the growth and development, metabolism, reproduction, and other physiological activities of insects, but its specific mechanism of action is still unclear. The aim of this study was to explore the relationship between expression of ERR and Vitellogenins (Vg) and the juvenile hormone (JH) and insulin/insulin-like growth factor/target of rapamycin (IIS/TOR) signaling pathways in Polyrhachis vicina Roger. P. vicina was used as the experimental model to clone the PvVg gene, perform double-stranded RNA synthesis and delivery and observe the effects of pharmacological treatments. The full-length PvVg cDNA product is 5586 bp. Higher PvVg mRNA expression was seen in the pupa and adults, and varying levels were seen in the different body parts of three different castes. RNA interference of PvVg expression led to disturbed development, an abnormal phenotype, and high mortality. PvVg RNAi also led to a reduction in mRNA levels of PvERR, ultraspiracle (PvUSP), forkhead box protein O (PvFOXO) and PvTOR genes in fourth instar larval, but a significant increase was seen in pupa and females. No significant change was seen in workers and males. After PvVg knockdown, application of exogenous JHIII reduced the expression of these genes in pupa and females, increased expression in workers, and decreased PvUSP mRNA expression in males. Both protein and mRNA expression levels of PvFOXO were affected by PvVg RNAi. PvERR RNAi increased PvVg expression in pupa and females and Kruppel-homolog 1 (PvKr-h1) and PvFOXO expression in males. The results of this study suggest that there is an interaction between PvERR and PvVg, and that crosstalk with the JH and IIS/TOR signaling pathways can affect development and reproduction. This effect is caste and developmental stage specific. We also speculate that the FOXO/USP complex participates in JH regulation of PvVg in P. vicina.
Identification of the major allergenic proteins from silkworm moth (Bombyx mori) involved in respiratory allergic diseases
2020, Allergologia et Immunopathologia
Citation Excerpt :
Vitelline is the most abundant protein in insect eggs and its precursor, vitellogenin is important during the reproductive cycle of various insect species. Yano et al. described that protein as an antibacterial agent from Bm pupa.21 Whilst in our study, the 45 kDa protein identified in moth-sensitized participants has an allergenic potential.
Moths are a significant source of indoor and outdoor aeroallergens. High prevalence of IgE-mediated sensitization was demonstrated in a group of patients with allergic respiratory diseases. There are no studies on adult stage of these moth species allergens involved in allergic respiratory reactions - the aim of this study.
36 participants were included in an experimental study, submitted to skin prick test with Bombyx mori wing extract and six other common allergens, as well as Western blot analysis with incubated nitrocellulose membrane impregnated with silkworm moth extract and human IgE-antibody. The participants were divided into 3 groups: 1) 21 allergic patients whose skin prick test was positive to Bombyx mori wing extract, 2) eight allergic patients whose skin prick test was positive to mite and negative to Bombyx mori extract 3) seven negative non-allergic subjects.
Among the 21 participants from group 1, 19 serum samples reacted to Bombyx mori extract by Western blot. All of them reacted to a protein at 80 kDa and five other proteins (66, 50, 45, 37 and 30 kDa) were identified in more than 50% of the individuals tested, considered as major allergenic proteins. Sera from seven out of eight patients sensitized to house dust mite demonstrated IgE-reactivity to Bombyx mori extract by Western blot analysis. Serum samples from healthy participants did not react at all.
Six major reactive proteins by immunoblot analysis from moth’s wings sensitized patients can be potential allergens. The one at 80 kDa is the major protein, seen in all IgE-reactive patients from group 1 and in none from group 2, yet to be identified. Future studies should be conducted to better characterize these proteins.
Molecular characterization and expression of vitellogenin gene from Spodoptera exigua exposed to cadmium stress
2016, Gene
Citation Excerpt :
Our finding suggested SEVg expression is associated with vitellogenesis in a tissue-, stage- and sex-dependent manner. This expression pattern is consistent with that of Vg in S. litura, but differs from B. mori and L. dispar in which Vg mRNA are first expressed at the laval stage (Shu et al., 2009; Yano et al., 1994a, 1994b; Hiremath and Lehtoma, 1997). The SEVg mRNA level changes with ovary development.
In this study, we cloned a full-length cDNA encoding vitellogenin (Vg) in Spodoptera exigua. The complete Vg cDNA consists of 5694 nucleotides with a long open reading frame encoding 1761 amino acid residues. The deduced amino acid sequence shared high similarity with the Vgs of other lepidopteran insects, particularly in the C-terminal region including the GL/ICG motif, five cysteine residues and DGXR motif. We analyzed the spatiotemporal expression of Vg transcripts in S. exigua by RT-PCR and Real-time Quantitative PCR. The results revealed that Vg was expressed specifically in the female fat body and was detectable after 5th day female pupae. The maximum level of Vg mRNA appeared in 48-h-old female adults and started to decrease quickly. When exposed to cadmium (Cd) (25–100 mg/kg), there was a significantly decreased Vg level in female adults. The information presented in this study suggested the Vg in S. exigua could be significantly affected by Cd pressure.
Molecular characterization and expression pattern of Spodoptera litura (Lepidoptera: Noctuidae) vitellogenin, and its response to lead stress
2009, Journal of Insect Physiology
Vitellogenin (Vg) cDNA from Spodoptera litura Fabricius was cloned and sequenced. The open reading frame (ORF) of Vg cDNA was 5247 nucleotides in length (GenBank Accession no. EU095334), which encoded for a protein of 1748 amino acids. S. litura Vg comprised three conserved regions (Vitellogenin-N domain, DUF1943 and von Willebrand factor type D domain (VWD)), a 17 amino-acid signal peptide and a RXXR cleavage signal (RTIR). The highly conserved GL/ICG motif, the DGXR motif and cysteine residues were found in the C-terminus of the Vg. Vg mRNA was found specifically in the female fat body. Vg expression was first transcribed in 6th day female pupae and levels increased with insect development. The maximum level of Vg mRNA appeared in 24-h-old adults. When S. litura larvae were exposed to lead (Pb) (25–200 mg Pb/kg), there was a significant inhibition in Vg of female adults. The start of Vg expression was advanced ahead by Pb, from 6th day pupae to 3rd day or 4th day pupae. Low levels of Vg in male adults were also induced by low concentrations of Pb (12.5 and 25 mg Pb/kg). These data show that Pb stress elicits an important Vg response in S. litura.
Molecular characteristics of insect vitellogenins
2008, Journal of Insect Physiology
Vitellogenins (Vgs) are precursors of the major egg storage protein, vitellin (Vn), in many oviparous animals. Insects Vgs are large molecules (∼200-kD) synthesized in the fat body in a process that involves substantial structural modifications (e.g., glycosylation, lipidation, phosphorylation, and proteolytic cleavage, etc.) of the nascent protein prior to its secretion and transport to the ovaries. However, the extent to which Vgs are processed in the fat body varies greatly among different insect groups. We provide evidence by cloning and peptide mapping of four Vg molecules from two cockroach species (Periplaneta americana and Leucophaea maderae) that, in hemimetabolous insects, the pro-Vg is cleaved into several polypeptides (ranging from 50-to 180-kD), unlike the holometabolans where the Vg precursor is cleaved into two polypeptides (one large and one small). An exception is the Vg of Apocrita (higher Hymenoptera) where the Vg gene product remains uncleaved. The yolk proteins (YPs) of higher Diptera (such as Drosophila) form a different family of proteins and are also not cleaved. So far, Vgs have been sequenced from 25 insect species; 9 of them belong to Hemimetabola and 16 to Holometabola. Alignment of the coding sequences revealed that some features, like the GL/ICG motif, cysteine residues, and a DGXR motif upstream of the GLI/CG motif, were highly conserved near the carboxy terminal of all insect Vgs. Moreover, a consensus RXXR cleavage sequence motif exists at the N-terminus of all sequences outside the Apocrita except for Lymantria dispar where it exists at the C-terminus. Phylogenetic analysis using 31 Vg sequences from 25 insect species reflects, in general, the current phylogenies of insects, suggesting that Vgs are still phylogenetically bound, although a divergence exists among them.

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¹: Present address: Pharmaceutical Research Center, Meiji Seika Kaisha, Ltd., Yokohama 222, Japan.

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Regular paperStructure and expression of mRNA for vitellogenin in Bombyx mori

Abstract

J. Insect Physiol.

Insect Biochem.

Dev. Biol.

Dev. Biol.

Arch. Biochem. Biophys.

Dev. Biol.

J. Biol. Chem.

J. Biol. Chem.

Dev. Biol.

Insect Biochem.

Insect Biochem.

Insect Biochem.

J. Biol. Chem.

J. Lipid Res.

Methods Enzymol.

Anal. Biochem.

Insect Biochem.

Gene

Cell

J. Mol. Biol.

Gene

J. Mol. Biol.

Biochem. Biophys. Res. Commun.

Insect Biochem.

Biochim. Biophys. Acta

Comp. Biochem. Physiol.

Regular paper
Structure and expression of mRNA for vitellogenin in Bombyx mori