Molecular cloning, expression and characterization of a novel vacuolar protein sorting 4 gene in silkworm, Bombyx mori

Xia, Hengchuan; Zhang, Chunxia; Fěng, Fang; Yuan, Yi; Zhou, Yang; Liu, Xiaoyong; Zhu, Kehua; Yao, Qin; Chen, Keping

doi:10.1007/s11033-012-1911-6

Cited by 4 publications

(3 citation statements)

References 26 publications

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“…In addition, mammalian Dlg can rescue the phenotype of a Drosophila Dlg mutant [11]. After the complete genome of B. mori was sequenced [12], the gene expression and regulation was widely analyzed [13,14,15]. However, there are no reports about the genes encoding the Scribble protein complex; including Scribble ( Scrib ), Discs large ( Dlg ) and Lethal giant larvae ( Lgl ) are found in silkworm, B. mori .…”

Section: Introductionmentioning

confidence: 99%

Molecular Expression of the Scribble Complex Genes, Dlg, Scrib and Lgl, in Silkworm, Bombyx mori

Liu

et al. 2013

Genes

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The Scribble protein complex genes, consisting of lethal giant larvae (Lgl), discs large (Dlg) and scribble (Scrib) genes, are components of an evolutionarily conserved genetic pathway that links the cell polarity in cells of humans and Drosophila. The tissue expression and developmental changes of the Scribble protein complex genes were documented using qRT-RCR method. The Lgl and Scrib genes could be detected in all the experimental tissues, including fat body, midgut, testis/ovary, wingdisc, trachea, malpighian tubule, hemolymph, prothoracic gland and silk gland. The Dlg gene, mainly expressed only in testis/ovary, could not be detected in prothoracic gland and hemolymph. In fat body, there were two higher expression stages of the three genes. The highest peak of the expression of the Lgl and Scrib genes in wingdisc lay at the 1st day of the 5th instar, but the Dlg gene was at 3rd day of 5th instar. The above results indicate that Scribble complex genes are involved in the process of molting and development of the wingdisc in the silkworm. This will be useful in the future for the elucidation of the detailed biological function of the three genes Scrib, Dlg and Lgl in B. mori.

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Section: Introductionmentioning

confidence: 99%

Molecular Expression of the Scribble Complex Genes, Dlg, Scrib and Lgl, in Silkworm, Bombyx mori

Liu

et al. 2013

Genes

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“…The silkworm ( Bombyx mori ) is of economic value and also a research model for lepidopteran insects. To explore the silkworm MVB pathway, we previously identified BmVps4 and BmVta1 as silkworm homologues for Vps4 and Vta1, respectively (Xia et al , , ). In the present study, we globally characterized the transcription and protein expression profiles of BmVps4 and BmVta1 using fluorescence quantitative real‐time PCR (qRT‐PCR) and Western blots.…”

Section: Introductionmentioning

confidence: 99%

Vacuolar protein sorting 4 is required for silkworm metamorphosis

Xia

Chen

Shao

et al. 2019

Insect Molecular Biology

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Vacuolar protein sorting 4 (Vps4) not only functions with its positive regulator vacuolar protein sorting 20‐associated 1 (Vta1) in the multivesicular body (MVB) pathway but also participates alone in MVB‐unrelated cellular processes. However, its physiological roles at the organism level remain rarely explored. We previously identified their respective homologues Bombyx mori Vps4 (BmVps4) and BmVta1 from the silkworm, a model organism for insect research. In this study, we performed fluorescence quantitative real‐time PCR and Western blot to globally characterize the transcription and protein expression profiles of BmVps4 and BmVta1 during silkworm development and in different silkworm tissues and organs. The results showed that they were significantly up‐regulated in metamorphosis, adulthood and embryogenesis relative to larval stages, and displayed a roughly similar tissue‐and‐organ specificity for transcriptions in silkworm larvae. Importantly, BmVps4 was down‐regulated during the early period of the fifth instar, reaching the lowest level of transcription on Day 6, then up‐regulated from Day 7 to the wandering, spinning and pupal stages, and down‐regulated again in adulthood. Moreover, knocking down BmVps4 by RNA interference significantly inhibited silk gland growth, shortened spinning time, prolonged pupation, reduced pupal size and weight, and increased moth wing defects. Together, our data demonstrate the critical and broad requirements for BmVps4 in silkworm metamorphosis.

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“…Silkworm ( Bombyx mori ) is an economic insect and a research model for Lepidoptera insects, and has also been used as a bioreactor to produce valuable bio-molecules. We previously identified a silkworm homologue for Vps4, named as BmVps4 ( Xia et al 2012 ). To expand our study of silkworm MVB pathway, in this report, we identified and cloned the silkworm homologous gene for VTA1 , expressed and characterized the protein designated as BmVta1.…”

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Identification and Characterization of BmVta1, a Bombyx mori (Lepidoptera: Bombycidae) Homologue for Vta1 That is Up-Regulated in Development

Xia

Shao

Liu

et al. 2017

Journal of Insect Science

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Vps20-associated 1 (Vta1) positively regulates Vacuolar protein sorting 4 (Vps4) to disassemble endosomal sorting complex required for transport III (ESCRT-III) for repeated uses in multivesicular body (MVB) pathway, virus budding and other processes. Currently, these proteins have mainly been studied in yeast and mammalian cells, while identities of them in insects remain largely unknown. We previously identified BmVps4, a Vps4 homologue from Bombyx mori. Here, we report the identification of a homologue for Vta1, designated as BmVta1. The BmVta1 cDNA contains an open reading frame of 933 bp and encodes a protein of 311 amino acid residues. We cloned BmVta1, expressed it in Escherichia coli, and prepared mouse polyclonal antibodies. Like BmVps4, BmVta1 is well conserved as shown by sequence analysis. Both proteins are localized in cytoplasm as revealed by subcellular location analysis. Interestingly, as revealed by semi-quantitative reverse transcription polymerase chain reaction (sqRT-PCR), transcriptions of BmVta1 and BmVps4 are highly up-regulated during silkworm metamorphosis and embryogenesis but down-regulated during larva stages, and are of higher levels in head, silk gland and testis than in Malpighian tube, fat body and ganglion, indicating important and similar roles of them in silkworm development and in silkworm tissues and organs. However, compared to BmVps4, the transcription of BmVta1 changes less drastically during development and is of much higher levels in midgut, ovary and hemolymph, suggesting the existence of distinct requirements of them in silkworm development and in certain tissues and organs.

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Molecular cloning, expression and characterization of a novel vacuolar protein sorting 4 gene in silkworm, Bombyx mori

Cited by 4 publications

References 26 publications

Molecular Expression of the Scribble Complex Genes, Dlg, Scrib and Lgl, in Silkworm, Bombyx mori

Molecular Expression of the Scribble Complex Genes, Dlg, Scrib and Lgl, in Silkworm, Bombyx mori

Vacuolar protein sorting 4 is required for silkworm metamorphosis

Identification and Characterization of BmVta1, a Bombyx mori (Lepidoptera: Bombycidae) Homologue for Vta1 That is Up-Regulated in Development

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