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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

Genomic structure and organization of BmTHYs.The top panel shows the genomic organization (A), the middle (B) and bottom (C) panels show the splicing patterns for BmTHY1 and BmTHY2, respectively. The introns are shown as lines and exons as boxes. Splicing sites are indicated by the diagonal dashed lines.
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pone.0140182.g003: Genomic structure and organization of BmTHYs.The top panel shows the genomic organization (A), the middle (B) and bottom (C) panels show the splicing patterns for BmTHY1 and BmTHY2, respectively. The introns are shown as lines and exons as boxes. Splicing sites are indicated by the diagonal dashed lines.

Mentions: To examine whether the two cloned cDNAs from BmTHYs are derived from the same gene, we cloned the corresponding genomic DNA sequence. By Genomic-F and Genomic-R, it could generate two bands using BmN cDNA as a template, while there exist only one product using the genomic as a template. Sequencing (S2 Fig) analysis revealed that this gene contained four exons: exon 1 (145bp), exon 2 (114bp), exon 3 (114bp) and exon 4 (140bp). BmTHY1 contains all the exon sequences, while BmTHY2 only includes exon1, 2 and 4 (Fig 3).


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)

Genomic structure and organization of BmTHYs.The top panel shows the genomic organization (A), the middle (B) and bottom (C) panels show the splicing patterns for BmTHY1 and BmTHY2, respectively. The introns are shown as lines and exons as boxes. Splicing sites are indicated by the diagonal dashed lines.
© Copyright Policy
Related In: Results  -  Collection

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

pone.0140182.g003: Genomic structure and organization of BmTHYs.The top panel shows the genomic organization (A), the middle (B) and bottom (C) panels show the splicing patterns for BmTHY1 and BmTHY2, respectively. The introns are shown as lines and exons as boxes. Splicing sites are indicated by the diagonal dashed lines.
Mentions: To examine whether the two cloned cDNAs from BmTHYs are derived from the same gene, we cloned the corresponding genomic DNA sequence. By Genomic-F and Genomic-R, it could generate two bands using BmN cDNA as a template, while there exist only one product using the genomic as a template. Sequencing (S2 Fig) analysis revealed that this gene contained four exons: exon 1 (145bp), exon 2 (114bp), exon 3 (114bp) and exon 4 (140bp). BmTHY1 contains all the exon sequences, while BmTHY2 only includes exon1, 2 and 4 (Fig 3).

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