Identification of nucleolar localization signal sequence of tumor metastasis suppressor gene-1 / 中华病理学杂志
Chinese Journal of Pathology
; (12): 767-771, 2011.
Article
em Zh
| WPRIM
| ID: wpr-358242
Biblioteca responsável:
WPRO
ABSTRACT
<p><b>OBJECTIVE</b>To identify the putative specific localization signal sequence of tumor metastasis suppressor gene-1 (TMSG-1) and to explore the mechanism of subcellular localization of TMSG-1 protein.</p><p><b>METHODS</b>Vectors expressing green fluorescence protein (GFP) tagged different TMSG-1 fragments were generated and transfected into human embryo kidney 293 (HEK293) cells. The expression of those fusion proteins was detected by Western blotting and their subcellular localizations were observed by laser confocal microscope.</p><p><b>RESULTS</b>GFP was fused with the native TMSG-1(aa1-380) or different fragments including T1 (aa1-70), T2 (aa1-128), T3 (aa129-380), T4 (aa71-128), T5 (aa71-179) and T6 (aa71-380). Anti-GFP Western blotting showed that these fusion proteins were successfully expressed. Under laser confocal microscope, GFP fused with fragment T4 (aa71-128) localized mainly in the nucleolus; GFP fused with fragment T6 (aa71-380) localized diffusely in the nucleus; while other fusion proteins with TMSG-1 (aa1-380) or fragment T1 (aa1-70), T2 (aa1-128), T3 (aa129-380) and T5 (aa71-179) localized in the cytoplasm. Fragment T4(Δ119-128) was generated from T4 with deletion of 10 amino acid of the C terminal. GFP fused with fragment T4(Δ119-128) remained in the nucleus, but no longer in the nucleolus.</p><p><b>CONCLUSIONS</b>There is a nucleolar localization signal (aa119-128 RRRRNQDRPS) within TMSG-1. This finding may have laid the foundation for further investigations into subcellular localization and function of TMSG-1.</p>
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Índice:
WPRIM
Assunto principal:
Plasmídeos
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Proteínas Recombinantes de Fusão
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Transfecção
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Nucléolo Celular
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Núcleo Celular
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Western Blotting
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Sequência de Aminoácidos
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Microscopia Confocal
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Sinais de Localização Nuclear
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Citoplasma
Tipo de estudo:
Diagnostic_studies
/
Prognostic_studies
Limite:
Humans
Idioma:
Zh
Revista:
Chinese Journal of Pathology
Ano de publicação:
2011
Tipo de documento:
Article