Identification of Ser2 proteins as major sericin components in the non-cocoon silk of Bombyx mori

Takasu, Yoko; Hata, Tamako; Uchino, Keiro; Zhang, Qiang

doi:10.1016/j.ibmb.2010.02.010

Cited by 87 publications

(95 citation statements)

References 18 publications

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“…Hence, the reduced weight of the cocoon shells made by w1-pnd P1A269/P1A269 larvae was considered most likely due to reduced fibroin content. SDS/PAGE in a gradient gel (5-20%) followed by immunoblotting with a FibL-specific antibody (13) showed that FibL and FibH proteins were present in the cocoon shells of w1-pnd +/+ larvae but not in the cocoons shells of w1-pnd P1A269/P1A269 larvae, in which only nonfibroin proteins, mainly sericins (15,(19)(20)(21)(22), were detectable (sericin 1-4; Fig. 2C).…”

Section: Expression and Physiological Effects Of Truncated P1a In Silmentioning

confidence: 99%

Bioengineered silkworms with butterfly cytotoxin-modified silk glands produce sericin cocoons with a utility for a new biomaterial

Otsuki

Yamamoto

Matsumoto

et al. 2017

Proc. Natl. Acad. Sci. U.S.A.

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Significance Specific gene functions have been successfully suppressed by gene silencing or editing in many organisms. However, genetic manipulation to suppress the function of a target tissue has not been achieved using cytotoxin genes. We established transgenic silkworms with posterior silk glands (PSGs) that express the enzymatic domain of the cytotoxin pierisin-1A (P1A). The larvae with the modified PSGs produced the sericin cocoons with potential utilities in tissue engineering. The targeted P1A expression was found to cause site-specific repression of certain protein synthesis that appeared to have no impact on the developmental stages of individuals. Thus, the new approach through targeted P1A expression could be applicable to the development of biologically useful model organisms with tissue-specific dysfunction.

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Section: Expression and Physiological Effects Of Truncated P1a In Silmentioning

confidence: 99%

Bioengineered silkworms with butterfly cytotoxin-modified silk glands produce sericin cocoons with a utility for a new biomaterial

Otsuki

Yamamoto

Matsumoto

et al. 2017

Proc. Natl. Acad. Sci. U.S.A.

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“…In addition to ser1, B. mori has two sericin genes (ser2 and ser3) that are expressed specifically in MSG-A and/or MSG-M (Fig. 1B) (35,36). As a first step to identify novel Antp target genes, we investigated whether ser2 and/or ser3 could be induced by Antp.…”

Section: Expression Of Sericin Genes Following Antp Misexpressionantpmentioning

confidence: 99%

A Hox Gene, Antennapedia, Regulates Expression of Multiple Major Silk Protein Genes in the Silkworm Bombyx mori

Tsubota

Tomita

Uchino

et al. 2016

Journal of Biological Chemistry

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Hox genes play a pivotal role in the determination of anteroposterior axis specificity during bilaterian animal development. They do so by acting as a master control and regulating the expression of genes important for development. Recently, however, we showed that Hox genes can also function in terminally differentiated tissue of the lepidopteran Bombyx mori. In this species, Antennapedia (Antp) regulates expression of sericin-1, a major silk protein gene, in the silk gland. Here, we investigated whether Antp can regulate expression of multiple genes in this tissue. By means of proteomic, RT-PCR, and in situ hybridization analyses, we demonstrate that misexpression of Antp in the posterior silk gland induced ectopic expression of major silk protein genes such as sericin-3, fhxh4, and fhxh5. These genes are normally expressed specifically in the middle silk gland as is Antp. Therefore, the evidence strongly suggests that Antp activates these silk protein genes in the middle silk gland. The putative sericin-1 activator complex (middle silk gland-intermoltspecific complex) can bind to the upstream regions of these genes, suggesting that Antp directly activates their expression. We also found that the pattern of gene expression was well conserved between B. mori and the wild species Bombyx mandarina, indicating that the gene regulation mechanism identified here is an evolutionarily conserved mechanism and not an artifact of the domestication of B. mori. We suggest that Hox genes have a role as a master control in terminally differentiated tissues, possibly acting as a primary regulator for a range of physiological processes.

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“…After 30-min incubation on ice, the homogenates were centrifuged at 12 000 × g for 15 min at 4ЊC and the supernatants were collected for subsequent SDS-PAGE and western blotting analysis. The crude Ser2 protein extract was isolated based on previous methods [44]. Briefly, the crude Ser2 proteins in the lumen of the anterior middle silk gland (MSG) were solubilized by immersing a roughly homogenized anterior MSG in water for 10 min.…”

Section: Protein Extraction and Detection Of Kac Proteins By Western mentioning

confidence: 99%

“…Furthermore, a large amount of Kac proteins containing various domains, such as acyl-CoA dehydrogenase domains, lipid transport protein domains, hemocyanin domains, and major domains of pyridoxal phosphate-dependent transferase, were found to be highly significantly enriched (p < 0.01) (Supporting Information Table 3). Acyl-CoA dehydrogenase and lipid transport proteins were the critical proteins involved in lipid metabolism and transport; hemocyanin domains exist in SPs, which are used to store amino acids as sources of amino acids during subsequent developmental stages [50]; pyridoxal phosphate-dependent transferase is primarily involved in the biosynthesis of amino acids and amino acid-derived metabolites [44]. These data strongly suggest that the cellular process of lipid metabolism and transport and biosynthesis, storage, and metabolism of amino acids may be strictly regulated by PTM lysine acetylation in silkworm.…”

Section: Functional Characteristics Of Kac Proteins In the Silkwormmentioning

confidence: 99%

Comprehensive profiling of lysine acetylation suggests the widespread function is regulated by protein acetylation in the silkworm, Bombyx mori

et al. 2015

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Lysine acetylation in proteins is a dynamic and reversible PTM and plays an important role in diverse cellular processes. In this study, using lysine-acetylation (Kac) peptide enrichment coupled with nano HPLC/MS/MS, we initially identified the acetylome in the silkworms. Overall, a total of 342 acetylated proteins with 667 Kac sites were identified in silkworm. Sequence motifs analysis around Kac sites revealed an enrichment of Y, F, and H in the +1 position, and F was also enriched in the +2 and -2 positions, indicating the presences of preferred amino acids around Kac sites in the silkworm. Functional analysis showed the acetylated proteins were primarily involved in some specific biological processes. Furthermore, lots of nutrient-storage proteins, such as apolipophorin, vitellogenin, storage proteins, and 30 K proteins, were highly acetylated, indicating lysine acetylation may represent a common regulatory mechanism of nutrient utilization in the silkworm. Interestingly, Ser2 proteins, the coating proteins of larval silk, were found to contain many Kac sites, suggesting lysine acetylation may be involved in the regulation of larval silk synthesis. This study is the first to identify the acetylome in a lepidoptera insect, and expands greatly the catalog of lysine acetylation substrates and sites in insects.

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Identification of Ser2 proteins as major sericin components in the non-cocoon silk of Bombyx mori

Cited by 87 publications

References 18 publications

Bioengineered silkworms with butterfly cytotoxin-modified silk glands produce sericin cocoons with a utility for a new biomaterial

Bioengineered silkworms with butterfly cytotoxin-modified silk glands produce sericin cocoons with a utility for a new biomaterial

A Hox Gene, Antennapedia, Regulates Expression of Multiple Major Silk Protein Genes in the Silkworm Bombyx mori

Comprehensive profiling of lysine acetylation suggests the widespread function is regulated by protein acetylation in the silkworm, Bombyx mori

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