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Functional differences between wild-type and mutant-type BRCA1-associated protein 1 tumor suppressor against malignant mesothelioma cells.

Hakiri S, Osada H, Ishiguro F, Murakami H, Murakami-Tonami Y, Yokoi K, Sekido Y - Cancer Sci. (2015)

Bottom Line: Transduction of the WT BAP1 vector into MM cells with homozygous deletion at the BAP1 3' side resulted in both inhibition of cell proliferation and anchorage-independent cell growth, whereas BAP1 mutants of a missense or C-terminal truncated form showed impaired growth inhibitory effects.Furthermore, using the MM cells with BAP1 deletion, we found that WT BAP1, and even a missense mutant, conferred a higher survival rate after IR compared to the control vector.Our results suggested that, whereas WT BAP1 suppresses MM cell proliferation and restores cell survival after IR damage, some mutant BAP1 may also moderately retain these functions.

View Article: PubMed Central - PubMed

Affiliation: Division of Molecular Oncology, Aichi Cancer Center Research Institute, Nagoya, Japan.

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Effects of BRCA1-associated protein 1 (BAP1) mutation on its own nuclear localization. (a) Endogenous WT BAP1 in NCI-H290 showed nuclear localization, whereas mutant BAP1 (p.F679LfsX37) in Y-MESO-9 showed cytoplasmic translocation. (b) Protein expression of exogenously transduced WT or mutant BAP1 (A95D, Y724X, and F679LfsX37) vectors into the malignant mesothelioma cell line with BAP1 deletion (Y-MESO-25). (c) Immunofluorescence analysis of subcellular BAP1 localization with exogenously transduced BAP1 vectors into the Y-MESO-25 cells. (d) Percentages of subcellular localization (c) were calculated. Wild-type BAP1 was mainly localized in the nucleus (N), whereas BAP1 mutants showed translocation in the cytoplasm (C). HD, homozygous deletion.
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fig02: Effects of BRCA1-associated protein 1 (BAP1) mutation on its own nuclear localization. (a) Endogenous WT BAP1 in NCI-H290 showed nuclear localization, whereas mutant BAP1 (p.F679LfsX37) in Y-MESO-9 showed cytoplasmic translocation. (b) Protein expression of exogenously transduced WT or mutant BAP1 (A95D, Y724X, and F679LfsX37) vectors into the malignant mesothelioma cell line with BAP1 deletion (Y-MESO-25). (c) Immunofluorescence analysis of subcellular BAP1 localization with exogenously transduced BAP1 vectors into the Y-MESO-25 cells. (d) Percentages of subcellular localization (c) were calculated. Wild-type BAP1 was mainly localized in the nucleus (N), whereas BAP1 mutants showed translocation in the cytoplasm (C). HD, homozygous deletion.

Mentions: BAP1 contains two NLS at the COOH-terminus, and is primarily localized in the nucleus.17 We used immunofluorescence analysis to determine whether or not the BAP1 mutations have an effect on its subcellular localization, because four mutations were predicted to result in the loss of the both NLS and one in the NLS2 at the COOH-terminus among the seven mutations. As expected, NCI-H290 cells, which harbor WT BAP1, showed endogenous BAP1 expression primarily in the nucleus (Fig.2a). In contrast, Y-MESO-9 cells, which lack the NLS2, showed endogenous BAP1 mainly in the cytoplasm. These results suggested that nuclear localization of endogenous BAP1 is impaired primarily by the loss of NLS, which was consistent with a previous study.30


Functional differences between wild-type and mutant-type BRCA1-associated protein 1 tumor suppressor against malignant mesothelioma cells.

Hakiri S, Osada H, Ishiguro F, Murakami H, Murakami-Tonami Y, Yokoi K, Sekido Y - Cancer Sci. (2015)

Effects of BRCA1-associated protein 1 (BAP1) mutation on its own nuclear localization. (a) Endogenous WT BAP1 in NCI-H290 showed nuclear localization, whereas mutant BAP1 (p.F679LfsX37) in Y-MESO-9 showed cytoplasmic translocation. (b) Protein expression of exogenously transduced WT or mutant BAP1 (A95D, Y724X, and F679LfsX37) vectors into the malignant mesothelioma cell line with BAP1 deletion (Y-MESO-25). (c) Immunofluorescence analysis of subcellular BAP1 localization with exogenously transduced BAP1 vectors into the Y-MESO-25 cells. (d) Percentages of subcellular localization (c) were calculated. Wild-type BAP1 was mainly localized in the nucleus (N), whereas BAP1 mutants showed translocation in the cytoplasm (C). HD, homozygous deletion.
© Copyright Policy - open-access
Related In: Results  -  Collection

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fig02: Effects of BRCA1-associated protein 1 (BAP1) mutation on its own nuclear localization. (a) Endogenous WT BAP1 in NCI-H290 showed nuclear localization, whereas mutant BAP1 (p.F679LfsX37) in Y-MESO-9 showed cytoplasmic translocation. (b) Protein expression of exogenously transduced WT or mutant BAP1 (A95D, Y724X, and F679LfsX37) vectors into the malignant mesothelioma cell line with BAP1 deletion (Y-MESO-25). (c) Immunofluorescence analysis of subcellular BAP1 localization with exogenously transduced BAP1 vectors into the Y-MESO-25 cells. (d) Percentages of subcellular localization (c) were calculated. Wild-type BAP1 was mainly localized in the nucleus (N), whereas BAP1 mutants showed translocation in the cytoplasm (C). HD, homozygous deletion.
Mentions: BAP1 contains two NLS at the COOH-terminus, and is primarily localized in the nucleus.17 We used immunofluorescence analysis to determine whether or not the BAP1 mutations have an effect on its subcellular localization, because four mutations were predicted to result in the loss of the both NLS and one in the NLS2 at the COOH-terminus among the seven mutations. As expected, NCI-H290 cells, which harbor WT BAP1, showed endogenous BAP1 expression primarily in the nucleus (Fig.2a). In contrast, Y-MESO-9 cells, which lack the NLS2, showed endogenous BAP1 mainly in the cytoplasm. These results suggested that nuclear localization of endogenous BAP1 is impaired primarily by the loss of NLS, which was consistent with a previous study.30

Bottom Line: Transduction of the WT BAP1 vector into MM cells with homozygous deletion at the BAP1 3' side resulted in both inhibition of cell proliferation and anchorage-independent cell growth, whereas BAP1 mutants of a missense or C-terminal truncated form showed impaired growth inhibitory effects.Furthermore, using the MM cells with BAP1 deletion, we found that WT BAP1, and even a missense mutant, conferred a higher survival rate after IR compared to the control vector.Our results suggested that, whereas WT BAP1 suppresses MM cell proliferation and restores cell survival after IR damage, some mutant BAP1 may also moderately retain these functions.

View Article: PubMed Central - PubMed

Affiliation: Division of Molecular Oncology, Aichi Cancer Center Research Institute, Nagoya, Japan.

Show MeSH
Related in: MedlinePlus