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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

Phylogenetic analysis of BmTHYs.Phylogenetic tree of various β-thymosins. The tree has been arbitrarily rooted between a clade consisting of the monomeric forms in vertebrate. GenBank numbers are as follows: Caenorhabditis briggsae (XP_002645512.1), Caenorhabditis remane (XP_003100241.1), Caenorhabditis brenneri (EGT34446.1), Caenorhabditis elegans (NP_509430.1), Ascaris suum (ERG81070.1), Stegodyphus mimosarum (KFM79832.1), Scylla paramamosain (ACY66642.1), Eriocheir sinensis thymosin-repeated protein 1,2 (ACP19740.1, ACP19741.1), Culex quinquefasciatus (XP_001863288.1), Drosophila melanogaster (NP_525065.1), Papilio xuthus (BAM17917.1), Helicoverpa armigera thymosin isoform 1,2 (ADD21556.1, ADD21557.10), Danaus plexippus (EHJ77183.1), Bombyx mori thymosin isoform 1,2 (NP_001040486.1, NP_001103818.1), Capitella teleta (ELU01379.1), Hermissenda crassicornis CSP24, CSP29 (AAN08024.1, AAN08022.1), Paracentrotus lividus (CAD29144.1), Strongylocentrotus purpuratus (NP_999791.1), Nanorana yunnanensis (ABQ12776.1), Homo sapiens (NP_066932.1), Fukomys damarensis (KFO19279.1), Gallus gallus (NP_001001315.1).
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pone.0140182.g002: Phylogenetic analysis of BmTHYs.Phylogenetic tree of various β-thymosins. The tree has been arbitrarily rooted between a clade consisting of the monomeric forms in vertebrate. GenBank numbers are as follows: Caenorhabditis briggsae (XP_002645512.1), Caenorhabditis remane (XP_003100241.1), Caenorhabditis brenneri (EGT34446.1), Caenorhabditis elegans (NP_509430.1), Ascaris suum (ERG81070.1), Stegodyphus mimosarum (KFM79832.1), Scylla paramamosain (ACY66642.1), Eriocheir sinensis thymosin-repeated protein 1,2 (ACP19740.1, ACP19741.1), Culex quinquefasciatus (XP_001863288.1), Drosophila melanogaster (NP_525065.1), Papilio xuthus (BAM17917.1), Helicoverpa armigera thymosin isoform 1,2 (ADD21556.1, ADD21557.10), Danaus plexippus (EHJ77183.1), Bombyx mori thymosin isoform 1,2 (NP_001040486.1, NP_001103818.1), Capitella teleta (ELU01379.1), Hermissenda crassicornis CSP24, CSP29 (AAN08024.1, AAN08022.1), Paracentrotus lividus (CAD29144.1), Strongylocentrotus purpuratus (NP_999791.1), Nanorana yunnanensis (ABQ12776.1), Homo sapiens (NP_066932.1), Fukomys damarensis (KFO19279.1), Gallus gallus (NP_001001315.1).

Mentions: As shown by the following phylogenetic tree analysis (Fig 2), it is clear that multirepeat β-thymosin is not a synapomorphy of the ecdysozoa, it also exist in annelida and mollusca. Note that, the monomeric in echinodermata and the multirepeat form in annelida and mollusca are in a clade, suggesting that the two forms are orthologous, which is incompatible with previous study [26]. Since other ecdysozoa phyla are not documented, Our phylogenetic tree is consistent with the previous study based on 18S rRNA sequence of β-thymosin which suggested that arthropoda be closely related to nematode worms [27].


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)

Phylogenetic analysis of BmTHYs.Phylogenetic tree of various β-thymosins. The tree has been arbitrarily rooted between a clade consisting of the monomeric forms in vertebrate. GenBank numbers are as follows: Caenorhabditis briggsae (XP_002645512.1), Caenorhabditis remane (XP_003100241.1), Caenorhabditis brenneri (EGT34446.1), Caenorhabditis elegans (NP_509430.1), Ascaris suum (ERG81070.1), Stegodyphus mimosarum (KFM79832.1), Scylla paramamosain (ACY66642.1), Eriocheir sinensis thymosin-repeated protein 1,2 (ACP19740.1, ACP19741.1), Culex quinquefasciatus (XP_001863288.1), Drosophila melanogaster (NP_525065.1), Papilio xuthus (BAM17917.1), Helicoverpa armigera thymosin isoform 1,2 (ADD21556.1, ADD21557.10), Danaus plexippus (EHJ77183.1), Bombyx mori thymosin isoform 1,2 (NP_001040486.1, NP_001103818.1), Capitella teleta (ELU01379.1), Hermissenda crassicornis CSP24, CSP29 (AAN08024.1, AAN08022.1), Paracentrotus lividus (CAD29144.1), Strongylocentrotus purpuratus (NP_999791.1), Nanorana yunnanensis (ABQ12776.1), Homo sapiens (NP_066932.1), Fukomys damarensis (KFO19279.1), Gallus gallus (NP_001001315.1).
© Copyright Policy
Related In: Results  -  Collection

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

pone.0140182.g002: Phylogenetic analysis of BmTHYs.Phylogenetic tree of various β-thymosins. The tree has been arbitrarily rooted between a clade consisting of the monomeric forms in vertebrate. GenBank numbers are as follows: Caenorhabditis briggsae (XP_002645512.1), Caenorhabditis remane (XP_003100241.1), Caenorhabditis brenneri (EGT34446.1), Caenorhabditis elegans (NP_509430.1), Ascaris suum (ERG81070.1), Stegodyphus mimosarum (KFM79832.1), Scylla paramamosain (ACY66642.1), Eriocheir sinensis thymosin-repeated protein 1,2 (ACP19740.1, ACP19741.1), Culex quinquefasciatus (XP_001863288.1), Drosophila melanogaster (NP_525065.1), Papilio xuthus (BAM17917.1), Helicoverpa armigera thymosin isoform 1,2 (ADD21556.1, ADD21557.10), Danaus plexippus (EHJ77183.1), Bombyx mori thymosin isoform 1,2 (NP_001040486.1, NP_001103818.1), Capitella teleta (ELU01379.1), Hermissenda crassicornis CSP24, CSP29 (AAN08024.1, AAN08022.1), Paracentrotus lividus (CAD29144.1), Strongylocentrotus purpuratus (NP_999791.1), Nanorana yunnanensis (ABQ12776.1), Homo sapiens (NP_066932.1), Fukomys damarensis (KFO19279.1), Gallus gallus (NP_001001315.1).
Mentions: As shown by the following phylogenetic tree analysis (Fig 2), it is clear that multirepeat β-thymosin is not a synapomorphy of the ecdysozoa, it also exist in annelida and mollusca. Note that, the monomeric in echinodermata and the multirepeat form in annelida and mollusca are in a clade, suggesting that the two forms are orthologous, which is incompatible with previous study [26]. Since other ecdysozoa phyla are not documented, Our phylogenetic tree is consistent with the previous study based on 18S rRNA sequence of β-thymosin which suggested that arthropoda be closely related to nematode worms [27].

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