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PMID:22383992
| Citation |
Zhang, W, Zhang, C, Lv, Z, Fang, D, Wang, D, Nie, Z, Yu, W, Lan, H, Jiang, C and Zhang, Y (2012) Molecular characterization, tissue distribution, subcellular localization and actin-sequestering function of a thymosin protein from silkworm. PLoS ONE 7:e31040 |
|---|---|
| Abstract |
We identified a novel gene encoding a Bombyx mori thymosin (BmTHY) protein from a cDNA library of silkworm pupae, which has an open reading frame (ORF) of 399 bp encoding 132 amino acids. It was found by bioinformatics that BmTHY gene consisted of three exons and two introns and BmTHY was highly homologous to thymosin betas (Tβ). BmTHY has a conserved motif LKHTET with only one amino acid difference from LKKTET, which is involved in Tβ binding to actin. A His-tagged BmTHY fusion protein (rBmTHY) with a molecular weight of approximately 18.4 kDa was expressed and purified to homogeneity. The purified fusion protein was used to produce anti-rBmTHY polyclonal antibodies in a New Zealand rabbit. Subcellular localization revealed that BmTHY can be found in both Bm5 cell (a silkworm ovary cell line) nucleus and cytoplasm but is primarily located in the nucleus. Western blotting and real-time RT-PCR showed that during silkworm developmental stages, BmTHY expression levels are highest in moth, followed by instar larvae, and are lowest in pupa and egg. BmTHY mRNA was universally distributed in most of fifth-instar larvae tissues (except testis). However, BmTHY was expressed in the head, ovary and epidermis during the larvae stage. BmTHY formed complexes with actin monomer, inhibited actin polymerization and cross-linked to actin. All the results indicated BmTHY might be an actin-sequestering protein and participate in silkworm development. |
| Links |
PubMed PMC3284464 Online version:10.1371/journal.pone.0031040 |
| Keywords |
Actins/chemistry; Amino Acid Motifs; Amino Acid Sequence; Animals; Base Sequence; Bombyx/physiology; Computational Biology/methods; DNA, Complementary/metabolism; Escherichia coli/metabolism; Gene Library; Introns; Microscopy, Fluorescence/methods; Molecular Sequence Data; Molecular Weight; Plasmids/metabolism; Rabbits; Real-Time Polymerase Chain Reaction/methods; Recombinant Fusion Proteins/metabolism; Recombinant Proteins/metabolism; Sequence Homology, Amino Acid; Thymosin/biosynthesis; Thymosin/genetics; Tissue Distribution |
| edit table |
Significance
Annotations
| Gene product | Qualifier | GO Term | Evidence Code | with/from | Aspect | Extension | Notes | Status |
|---|---|---|---|---|---|---|---|---|
|
part_of |
GO:0005634: nucleus |
ECO:0000314: direct assay evidence used in manual assertion |
C |
Seeded From UniProt |
complete | |||
|
part_of |
GO:0005737: cytoplasm |
ECO:0000314: direct assay evidence used in manual assertion |
C |
Seeded From UniProt |
complete | |||
| GO:0005634: nucleus |
ECO:0000314: |
C |
Figure 10. |
complete | ||||
| GO:0005737: cytoplasm |
ECO:0000314: |
C |
Figure 10. |
complete | ||||
See also
References
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