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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

Sequence alignment of BmTHYs.Amino acids are colored according to the conservation (Black represents the most conservative, followed by dark gray, then the weakest is light gray) GenBank numbers are as follows: Bombyx mori thymosin isoform 1,2 (NP_001040486.1, NP_001103818.1), Helicoverpa armigera thymosin isoform 1,2 (ADD21556.1, ADD21557.10), Culex quinquefasciatus (XP_001863288.1), Drosophila melanogaster (NP_525065.1), Caenorhabditis elegans (NP_509430.1), Eriocheir sinensis thymosin-repeated protein 1 (ACP19740.1), Homo sapiens (NP_066932.1), Branchiostoma belcheri (AAK72482.1), Myotis brandtii (EPQ12153.1) Nanorana yunnanensis (ABQ12776.1), Amolops loloensis (ABG78789.1).
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pone.0140182.g001: Sequence alignment of BmTHYs.Amino acids are colored according to the conservation (Black represents the most conservative, followed by dark gray, then the weakest is light gray) GenBank numbers are as follows: Bombyx mori thymosin isoform 1,2 (NP_001040486.1, NP_001103818.1), Helicoverpa armigera thymosin isoform 1,2 (ADD21556.1, ADD21557.10), Culex quinquefasciatus (XP_001863288.1), Drosophila melanogaster (NP_525065.1), Caenorhabditis elegans (NP_509430.1), Eriocheir sinensis thymosin-repeated protein 1 (ACP19740.1), Homo sapiens (NP_066932.1), Branchiostoma belcheri (AAK72482.1), Myotis brandtii (EPQ12153.1) Nanorana yunnanensis (ABQ12776.1), Amolops loloensis (ABG78789.1).

Mentions: We amplified two specific bands using PCR method, named as BmTHY1 and BmTHY2. As revealed by sequencing and BLAST analysis, the coding regions of BmTHY2 are completely included in that of BmTHY1. Compared to protein sequences from other species, the two cloned BmTHYs protein sequences show a high identity with them, including HaTHYs from H. armigera, Cib B from D. melanogaster etc. Notably, only the second and the fourth THY domains are highly similar to vertebrate THY, the first THY domain is quite different with binding sites mutated to "LRDV" from" LKK/HT", and the third one is unique that exists only in the lepidoptera insects (Fig 1), suggesting the lepidoptera insects including silkworm and cotton bollworm may have specific roles for such an organization of β-thymosins. The binding motif of each THY domain for G-actin is quite different from "LKHT", which is very conservative in vertebrate. Besides, the silkworm β-thymosins also contain an elongated N-terminal sequence like other invertebrate β-thymosins, which is shown to be able to enhance the affinity of binding to G-actin compared to vertebrate β-thymosins [13].


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)

Sequence alignment of BmTHYs.Amino acids are colored according to the conservation (Black represents the most conservative, followed by dark gray, then the weakest is light gray) GenBank numbers are as follows: Bombyx mori thymosin isoform 1,2 (NP_001040486.1, NP_001103818.1), Helicoverpa armigera thymosin isoform 1,2 (ADD21556.1, ADD21557.10), Culex quinquefasciatus (XP_001863288.1), Drosophila melanogaster (NP_525065.1), Caenorhabditis elegans (NP_509430.1), Eriocheir sinensis thymosin-repeated protein 1 (ACP19740.1), Homo sapiens (NP_066932.1), Branchiostoma belcheri (AAK72482.1), Myotis brandtii (EPQ12153.1) Nanorana yunnanensis (ABQ12776.1), Amolops loloensis (ABG78789.1).
© Copyright Policy
Related In: Results  -  Collection

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

pone.0140182.g001: Sequence alignment of BmTHYs.Amino acids are colored according to the conservation (Black represents the most conservative, followed by dark gray, then the weakest is light gray) GenBank numbers are as follows: Bombyx mori thymosin isoform 1,2 (NP_001040486.1, NP_001103818.1), Helicoverpa armigera thymosin isoform 1,2 (ADD21556.1, ADD21557.10), Culex quinquefasciatus (XP_001863288.1), Drosophila melanogaster (NP_525065.1), Caenorhabditis elegans (NP_509430.1), Eriocheir sinensis thymosin-repeated protein 1 (ACP19740.1), Homo sapiens (NP_066932.1), Branchiostoma belcheri (AAK72482.1), Myotis brandtii (EPQ12153.1) Nanorana yunnanensis (ABQ12776.1), Amolops loloensis (ABG78789.1).
Mentions: We amplified two specific bands using PCR method, named as BmTHY1 and BmTHY2. As revealed by sequencing and BLAST analysis, the coding regions of BmTHY2 are completely included in that of BmTHY1. Compared to protein sequences from other species, the two cloned BmTHYs protein sequences show a high identity with them, including HaTHYs from H. armigera, Cib B from D. melanogaster etc. Notably, only the second and the fourth THY domains are highly similar to vertebrate THY, the first THY domain is quite different with binding sites mutated to "LRDV" from" LKK/HT", and the third one is unique that exists only in the lepidoptera insects (Fig 1), suggesting the lepidoptera insects including silkworm and cotton bollworm may have specific roles for such an organization of β-thymosins. The binding motif of each THY domain for G-actin is quite different from "LKHT", which is very conservative in vertebrate. Besides, the silkworm β-thymosins also contain an elongated N-terminal sequence like other invertebrate β-thymosins, which is shown to be able to enhance the affinity of binding to G-actin compared to vertebrate β-thymosins [13].

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