Limits...
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 protein-protein interaction analysis.The total protein of midgut was used for the GST pull down (A, B) and Co-IP (C, D) to detect proteins interacting with BmTHYs. The GST tagged BmTHY1 and BmTHY2 were purified and used to pull down actin (A) and 14-3-3 (B). The Co-IP was performed using anti-actin (C) and anti-14-3-3(D) antibodies as baits, and anti-BmTHY2 antiserum was used for Western blot analysis.
© Copyright Policy
Related In: Results  -  Collection

License
getmorefigures.php?uid=PMC4608725&req=5

pone.0140182.g006: The protein-protein interaction analysis.The total protein of midgut was used for the GST pull down (A, B) and Co-IP (C, D) to detect proteins interacting with BmTHYs. The GST tagged BmTHY1 and BmTHY2 were purified and used to pull down actin (A) and 14-3-3 (B). The Co-IP was performed using anti-actin (C) and anti-14-3-3(D) antibodies as baits, and anti-BmTHY2 antiserum was used for Western blot analysis.

Mentions: To explore which protein of midgut can interact with BmTHYs, GST pull down and Co-IP assay were carried out. The results showed that both of them can bind to 14-3-3. Unexpectedly, using the infected midgut as samples, we found both of BmTHYs can interact with Ubiquitin of BmNPV (Fig 6).


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 protein-protein interaction analysis.The total protein of midgut was used for the GST pull down (A, B) and Co-IP (C, D) to detect proteins interacting with BmTHYs. The GST tagged BmTHY1 and BmTHY2 were purified and used to pull down actin (A) and 14-3-3 (B). The Co-IP was performed using anti-actin (C) and anti-14-3-3(D) antibodies as baits, and anti-BmTHY2 antiserum was used for Western blot analysis.
© Copyright Policy
Related In: Results  -  Collection

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

pone.0140182.g006: The protein-protein interaction analysis.The total protein of midgut was used for the GST pull down (A, B) and Co-IP (C, D) to detect proteins interacting with BmTHYs. The GST tagged BmTHY1 and BmTHY2 were purified and used to pull down actin (A) and 14-3-3 (B). The Co-IP was performed using anti-actin (C) and anti-14-3-3(D) antibodies as baits, and anti-BmTHY2 antiserum was used for Western blot analysis.
Mentions: To explore which protein of midgut can interact with BmTHYs, GST pull down and Co-IP assay were carried out. The results showed that both of them can bind to 14-3-3. Unexpectedly, using the infected midgut as samples, we found both of BmTHYs can interact with Ubiquitin of BmNPV (Fig 6).

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