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Hsp10 nuclear localization and changes in lung cells response to cigarette smoke suggest novel roles for this chaperonin.

Corrao S, Anzalone R, Lo Iacono M, Corsello T, Di Stefano A, D'Anna SE, Balbi B, Carone M, Sala A, Corona D, Timperio AM, Zolla L, Farina F, de Macario EC, Macario AJ, Cappello F, La Rocca G - Open Biol (2014)

Bottom Line: Heat-shock protein (Hsp)10 is the co-chaperone for Hsp60 inside mitochondria, but it also resides outside the organelle.Variations in its levels and intracellular distribution have been documented in pathological conditions, e.g. cancer and chronic obstructive pulmonary disease (COPD).Noteworthy findings are: (i) Hsp10 occurred in nuclei of epithelial and lamina propria cells of bronchial mucosa from non-smokers and smokers; (ii) human bronchial epithelial (16HBE) and lung fibroblast (HFL-1) cells, in vitro, showed Hsp10 in the nucleus, before and after CSE exposure; (iii) CSE stimulation did not increase the levels of Hsp10 but did elicit qualitative changes as indicated by molecular weight and isoelectric point shifts; and (iv) Hsp10 nuclear levels increased after CSE stimulation in HFL-1, indicating cytosol to nucleus migration, and although Hsp10 did not bind DNA, it bound a DNA-associated protein.

View Article: PubMed Central - PubMed

Affiliation: Istituto Euro-Mediterraneo di Scienza e Tecnologia (IEMEST), Palermo, Italy Dipartimento di Biomedicina Sperimentale e Neuroscienze Cliniche, Università degli Studi di Palermo, Palermo, Italy.

ABSTRACT
Heat-shock protein (Hsp)10 is the co-chaperone for Hsp60 inside mitochondria, but it also resides outside the organelle. Variations in its levels and intracellular distribution have been documented in pathological conditions, e.g. cancer and chronic obstructive pulmonary disease (COPD). Here, we show that Hsp10 in COPD undergoes changes at the molecular and subcellular levels in bronchial cells from human specimens and derived cell lines, intact or subjected to stress induced by cigarette smoke extract (CSE). Noteworthy findings are: (i) Hsp10 occurred in nuclei of epithelial and lamina propria cells of bronchial mucosa from non-smokers and smokers; (ii) human bronchial epithelial (16HBE) and lung fibroblast (HFL-1) cells, in vitro, showed Hsp10 in the nucleus, before and after CSE exposure; (iii) CSE stimulation did not increase the levels of Hsp10 but did elicit qualitative changes as indicated by molecular weight and isoelectric point shifts; and (iv) Hsp10 nuclear levels increased after CSE stimulation in HFL-1, indicating cytosol to nucleus migration, and although Hsp10 did not bind DNA, it bound a DNA-associated protein.

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Immunocytochemistry for Hsp10 and Hsp60. (a) Levels of Hsp10 and Hsp60 in 16HBE and HFL-1 cells before (NT) and after (10%) incubation with CSE at 10%. No statistically significant differences were found between the groups (p-values ranging between 0.48 and 0.68). (b) Representative microphotographs showing immunolocalization by immunohistochemistry of Hsp10 and Hsp60 in 16HBE and HFL-1 cells. The insets indicate nuclear localization of Hsp10. Scale bars, 10 µm (figures) and 1 µm (insets).
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RSOB140125F3: Immunocytochemistry for Hsp10 and Hsp60. (a) Levels of Hsp10 and Hsp60 in 16HBE and HFL-1 cells before (NT) and after (10%) incubation with CSE at 10%. No statistically significant differences were found between the groups (p-values ranging between 0.48 and 0.68). (b) Representative microphotographs showing immunolocalization by immunohistochemistry of Hsp10 and Hsp60 in 16HBE and HFL-1 cells. The insets indicate nuclear localization of Hsp10. Scale bars, 10 µm (figures) and 1 µm (insets).

Mentions: We quantified the levels of Hsp10 and Hsp60 in 16HBE and HFL-1 cells before and after CSE treatment. Hsp10 and Hsp60 levels did not show quantitative changes after treatment (p > 0.05; figure 3a). Hsp60 was present in the cytoplasm, often with the granular aspect typical of mitochondrial localization (figure 3b). Hsp10 showed not only a wide cytoplasmic, granular distribution but also an appreciable positivity at the nuclear level (figure 3b, insets). These data were in agreement with the in vivo immunohistochemical results obtained with bronchial mucosa cells as described above, and showed nuclear localization of Hsp10 in treated and untreated cells. To the best of our knowledge, positivity for Hsp10 in the nucleus of airways mucosa cells has never been described before.Figure 3.


Hsp10 nuclear localization and changes in lung cells response to cigarette smoke suggest novel roles for this chaperonin.

Corrao S, Anzalone R, Lo Iacono M, Corsello T, Di Stefano A, D'Anna SE, Balbi B, Carone M, Sala A, Corona D, Timperio AM, Zolla L, Farina F, de Macario EC, Macario AJ, Cappello F, La Rocca G - Open Biol (2014)

Immunocytochemistry for Hsp10 and Hsp60. (a) Levels of Hsp10 and Hsp60 in 16HBE and HFL-1 cells before (NT) and after (10%) incubation with CSE at 10%. No statistically significant differences were found between the groups (p-values ranging between 0.48 and 0.68). (b) Representative microphotographs showing immunolocalization by immunohistochemistry of Hsp10 and Hsp60 in 16HBE and HFL-1 cells. The insets indicate nuclear localization of Hsp10. Scale bars, 10 µm (figures) and 1 µm (insets).
© Copyright Policy - open-access
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC4221893&req=5

RSOB140125F3: Immunocytochemistry for Hsp10 and Hsp60. (a) Levels of Hsp10 and Hsp60 in 16HBE and HFL-1 cells before (NT) and after (10%) incubation with CSE at 10%. No statistically significant differences were found between the groups (p-values ranging between 0.48 and 0.68). (b) Representative microphotographs showing immunolocalization by immunohistochemistry of Hsp10 and Hsp60 in 16HBE and HFL-1 cells. The insets indicate nuclear localization of Hsp10. Scale bars, 10 µm (figures) and 1 µm (insets).
Mentions: We quantified the levels of Hsp10 and Hsp60 in 16HBE and HFL-1 cells before and after CSE treatment. Hsp10 and Hsp60 levels did not show quantitative changes after treatment (p > 0.05; figure 3a). Hsp60 was present in the cytoplasm, often with the granular aspect typical of mitochondrial localization (figure 3b). Hsp10 showed not only a wide cytoplasmic, granular distribution but also an appreciable positivity at the nuclear level (figure 3b, insets). These data were in agreement with the in vivo immunohistochemical results obtained with bronchial mucosa cells as described above, and showed nuclear localization of Hsp10 in treated and untreated cells. To the best of our knowledge, positivity for Hsp10 in the nucleus of airways mucosa cells has never been described before.Figure 3.

Bottom Line: Heat-shock protein (Hsp)10 is the co-chaperone for Hsp60 inside mitochondria, but it also resides outside the organelle.Variations in its levels and intracellular distribution have been documented in pathological conditions, e.g. cancer and chronic obstructive pulmonary disease (COPD).Noteworthy findings are: (i) Hsp10 occurred in nuclei of epithelial and lamina propria cells of bronchial mucosa from non-smokers and smokers; (ii) human bronchial epithelial (16HBE) and lung fibroblast (HFL-1) cells, in vitro, showed Hsp10 in the nucleus, before and after CSE exposure; (iii) CSE stimulation did not increase the levels of Hsp10 but did elicit qualitative changes as indicated by molecular weight and isoelectric point shifts; and (iv) Hsp10 nuclear levels increased after CSE stimulation in HFL-1, indicating cytosol to nucleus migration, and although Hsp10 did not bind DNA, it bound a DNA-associated protein.

View Article: PubMed Central - PubMed

Affiliation: Istituto Euro-Mediterraneo di Scienza e Tecnologia (IEMEST), Palermo, Italy Dipartimento di Biomedicina Sperimentale e Neuroscienze Cliniche, Università degli Studi di Palermo, Palermo, Italy.

ABSTRACT
Heat-shock protein (Hsp)10 is the co-chaperone for Hsp60 inside mitochondria, but it also resides outside the organelle. Variations in its levels and intracellular distribution have been documented in pathological conditions, e.g. cancer and chronic obstructive pulmonary disease (COPD). Here, we show that Hsp10 in COPD undergoes changes at the molecular and subcellular levels in bronchial cells from human specimens and derived cell lines, intact or subjected to stress induced by cigarette smoke extract (CSE). Noteworthy findings are: (i) Hsp10 occurred in nuclei of epithelial and lamina propria cells of bronchial mucosa from non-smokers and smokers; (ii) human bronchial epithelial (16HBE) and lung fibroblast (HFL-1) cells, in vitro, showed Hsp10 in the nucleus, before and after CSE exposure; (iii) CSE stimulation did not increase the levels of Hsp10 but did elicit qualitative changes as indicated by molecular weight and isoelectric point shifts; and (iv) Hsp10 nuclear levels increased after CSE stimulation in HFL-1, indicating cytosol to nucleus migration, and although Hsp10 did not bind DNA, it bound a DNA-associated protein.

Show MeSH
Related in: MedlinePlus