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Molecular and Physiological Characterization of Two Novel Multirepeat β-Thymosins from Silkworm, Bombyx mori.

Ma S, Kang Z, Lü P, Yang Y, Yao Q, Xia H, Chen K - PLoS ONE (2015)

Bottom Line: Amazingly, the expression of BmTHY2 was hugely increased during the pupae stage, indicating a specialized role in this period.The expression of these proteins was gradually decreased in BmN cells infected by BmNPV, suggesting they may play different roles in the virus infection.In addition, both BmTHY1 and BmTHY2 can interact with 14-3-3 of silkworm and Ubiquitin of BmNPV as shown by GST pull down and Co-IP assays, consistent with their roles in the regulation of the development of nervous system.

View Article: PubMed Central - PubMed

Affiliation: Institute of Life Sciences, Jiangsu University, Zhenjiang, Jiangsu, P. R. China.

ABSTRACT
β-thymosin plays important roles in the development of the lymphatic system and the central nervous system in vertebrates. However, its role and function in invertebrates remain much less explored. Here, we firstly isolated a gene encoding β-thymosin in silkworm (Bombyx mori L.). Interestingly, this gene encodes two polypeptides, named as BmTHY1 and BmTHY2, via two different modes of RNA splicing. The recombinant proteins fused with an N-term GST tag were over-expressed in Escherichia coli (E. coli) and further purified to near homogenity to prepare mouse antibodies. The Western blot analysis showed that these proteins were expressed in various tissues and organs, as well as in different developmental stages. Amazingly, the expression of BmTHY2 was hugely increased during the pupae stage, indicating a specialized role in this period. The expression of these proteins was gradually decreased in BmN cells infected by BmNPV, suggesting they may play different roles in the virus infection. In addition, both BmTHY1 and BmTHY2 can interact with 14-3-3 of silkworm and Ubiquitin of BmNPV as shown by GST pull down and Co-IP assays, consistent with their roles in the regulation of the development of nervous system.

No MeSH data available.


Related in: MedlinePlus

The Spatiotemporal expression profiles of BmTHYs.Western blot analysis of the expression levels of BmTHYs in different developmental stage (A) and in different tissues and organs (B). The mass of each lane’s total protein were 50 μg, and the sample were equalized. (A) Lane 1: egg; Lane 2: 1st instar; Lane 3: 2nd instar; Lane 4: 3rd instar; Lane 5: 4th instar; Lane 6: 5th instar; Lane 7: pupae; Lane 8: moth. (B) Lane 1: midgut; Lane 2: testis; Lane 3: ovary; Lane 4: head; Lane 5: fat body; Lane 6: hemolymph.
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pone.0140182.g005: The Spatiotemporal expression profiles of BmTHYs.Western blot analysis of the expression levels of BmTHYs in different developmental stage (A) and in different tissues and organs (B). The mass of each lane’s total protein were 50 μg, and the sample were equalized. (A) Lane 1: egg; Lane 2: 1st instar; Lane 3: 2nd instar; Lane 4: 3rd instar; Lane 5: 4th instar; Lane 6: 5th instar; Lane 7: pupae; Lane 8: moth. (B) Lane 1: midgut; Lane 2: testis; Lane 3: ovary; Lane 4: head; Lane 5: fat body; Lane 6: hemolymph.

Mentions: Western blot was performed to determine BmTHYs expression levels. As shown in Fig 5A, the two β-thymosins were widely expressed in all the samples covering the whole life cycle of silkworms. Amazingly, during the pupae stage the BmTHY2 were greatly over-expressed, indicating it may be involved in metamorphosis.


Molecular and Physiological Characterization of Two Novel Multirepeat β-Thymosins from Silkworm, Bombyx mori.

Ma S, Kang Z, Lü P, Yang Y, Yao Q, Xia H, Chen K - PLoS ONE (2015)

The Spatiotemporal expression profiles of BmTHYs.Western blot analysis of the expression levels of BmTHYs in different developmental stage (A) and in different tissues and organs (B). The mass of each lane’s total protein were 50 μg, and the sample were equalized. (A) Lane 1: egg; Lane 2: 1st instar; Lane 3: 2nd instar; Lane 4: 3rd instar; Lane 5: 4th instar; Lane 6: 5th instar; Lane 7: pupae; Lane 8: moth. (B) Lane 1: midgut; Lane 2: testis; Lane 3: ovary; Lane 4: head; Lane 5: fat body; Lane 6: hemolymph.
© Copyright Policy
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC4608725&req=5

pone.0140182.g005: The Spatiotemporal expression profiles of BmTHYs.Western blot analysis of the expression levels of BmTHYs in different developmental stage (A) and in different tissues and organs (B). The mass of each lane’s total protein were 50 μg, and the sample were equalized. (A) Lane 1: egg; Lane 2: 1st instar; Lane 3: 2nd instar; Lane 4: 3rd instar; Lane 5: 4th instar; Lane 6: 5th instar; Lane 7: pupae; Lane 8: moth. (B) Lane 1: midgut; Lane 2: testis; Lane 3: ovary; Lane 4: head; Lane 5: fat body; Lane 6: hemolymph.
Mentions: Western blot was performed to determine BmTHYs expression levels. As shown in Fig 5A, the two β-thymosins were widely expressed in all the samples covering the whole life cycle of silkworms. Amazingly, during the pupae stage the BmTHY2 were greatly over-expressed, indicating it may be involved in metamorphosis.

Bottom Line: Amazingly, the expression of BmTHY2 was hugely increased during the pupae stage, indicating a specialized role in this period.The expression of these proteins was gradually decreased in BmN cells infected by BmNPV, suggesting they may play different roles in the virus infection.In addition, both BmTHY1 and BmTHY2 can interact with 14-3-3 of silkworm and Ubiquitin of BmNPV as shown by GST pull down and Co-IP assays, consistent with their roles in the regulation of the development of nervous system.

View Article: PubMed Central - PubMed

Affiliation: Institute of Life Sciences, Jiangsu University, Zhenjiang, Jiangsu, P. R. China.

ABSTRACT
β-thymosin plays important roles in the development of the lymphatic system and the central nervous system in vertebrates. However, its role and function in invertebrates remain much less explored. Here, we firstly isolated a gene encoding β-thymosin in silkworm (Bombyx mori L.). Interestingly, this gene encodes two polypeptides, named as BmTHY1 and BmTHY2, via two different modes of RNA splicing. The recombinant proteins fused with an N-term GST tag were over-expressed in Escherichia coli (E. coli) and further purified to near homogenity to prepare mouse antibodies. The Western blot analysis showed that these proteins were expressed in various tissues and organs, as well as in different developmental stages. Amazingly, the expression of BmTHY2 was hugely increased during the pupae stage, indicating a specialized role in this period. The expression of these proteins was gradually decreased in BmN cells infected by BmNPV, suggesting they may play different roles in the virus infection. In addition, both BmTHY1 and BmTHY2 can interact with 14-3-3 of silkworm and Ubiquitin of BmNPV as shown by GST pull down and Co-IP assays, consistent with their roles in the regulation of the development of nervous system.

No MeSH data available.


Related in: MedlinePlus