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Differential role of nonmuscle myosin II isoforms during blebbing of MCF-7 cells
Bleb formation has been correlated with nonmuscle myosin II (NM-II) activity. Whether three isoforms of NM-II (NM-IIA, -IIB and -IIC) have the same or differential roles in bleb formation is not well understood. Here we report that ectopically expressed, GFP-tagged NM-II isoforms exhibit different t...
Autores principales: | , , , , , , , , |
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Formato: | Online Artículo Texto |
Lenguaje: | English |
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The American Society for Cell Biology
2017
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Materias: | |
Acceso en línea: | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC5391180/ https://www.ncbi.nlm.nih.gov/pubmed/28251924 http://dx.doi.org/10.1091/mbc.E16-07-0524 |
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author | Dey, Sumit K. Singh, Raman K. Chattoraj, Shyamtanu Saha, Shekhar Das, Alakesh Bhattacharyya, Kankan Sengupta, Kaushik Sen, Shamik Jana, Siddhartha S. |
author_facet | Dey, Sumit K. Singh, Raman K. Chattoraj, Shyamtanu Saha, Shekhar Das, Alakesh Bhattacharyya, Kankan Sengupta, Kaushik Sen, Shamik Jana, Siddhartha S. |
author_sort | Dey, Sumit K. |
collection | PubMed |
description | Bleb formation has been correlated with nonmuscle myosin II (NM-II) activity. Whether three isoforms of NM-II (NM-IIA, -IIB and -IIC) have the same or differential roles in bleb formation is not well understood. Here we report that ectopically expressed, GFP-tagged NM-II isoforms exhibit different types of membrane protrusions, such as multiple blebs, lamellipodia, combinations of both, or absence of any such protrusions in MCF-7 cells. Quantification suggests that 50% of NM-IIA-GFP–, 29% of NM-IIB-GFP–, and 19% of NM-IIC1-GFP–expressing MCF-7 cells show multiple bleb formation, compared with 36% of cells expressing GFP alone. Of interest, NM-IIB has an almost 50% lower rate of dissociation from actin filament than NM-IIA and –IIC1 as determined by FRET analysis both at cell and bleb cortices. We induced bleb formation by disruption of the cortex and found that all three NM-II-GFP isoforms can reappear and form filaments but to different degrees in the growing bleb. NM-IIB-GFP can form filaments in blebs in 41% of NM-IIB-GFP–expressing cells, whereas filaments form in only 12 and 3% of cells expressing NM-IIA-GFP and NM-IIC1-GFP, respectively. These studies suggest that NM-II isoforms have differential roles in the bleb life cycle. |
format | Online Article Text |
id | pubmed-5391180 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2017 |
publisher | The American Society for Cell Biology |
record_format | MEDLINE/PubMed |
spelling | pubmed-53911802017-06-30 Differential role of nonmuscle myosin II isoforms during blebbing of MCF-7 cells Dey, Sumit K. Singh, Raman K. Chattoraj, Shyamtanu Saha, Shekhar Das, Alakesh Bhattacharyya, Kankan Sengupta, Kaushik Sen, Shamik Jana, Siddhartha S. Mol Biol Cell Articles Bleb formation has been correlated with nonmuscle myosin II (NM-II) activity. Whether three isoforms of NM-II (NM-IIA, -IIB and -IIC) have the same or differential roles in bleb formation is not well understood. Here we report that ectopically expressed, GFP-tagged NM-II isoforms exhibit different types of membrane protrusions, such as multiple blebs, lamellipodia, combinations of both, or absence of any such protrusions in MCF-7 cells. Quantification suggests that 50% of NM-IIA-GFP–, 29% of NM-IIB-GFP–, and 19% of NM-IIC1-GFP–expressing MCF-7 cells show multiple bleb formation, compared with 36% of cells expressing GFP alone. Of interest, NM-IIB has an almost 50% lower rate of dissociation from actin filament than NM-IIA and –IIC1 as determined by FRET analysis both at cell and bleb cortices. We induced bleb formation by disruption of the cortex and found that all three NM-II-GFP isoforms can reappear and form filaments but to different degrees in the growing bleb. NM-IIB-GFP can form filaments in blebs in 41% of NM-IIB-GFP–expressing cells, whereas filaments form in only 12 and 3% of cells expressing NM-IIA-GFP and NM-IIC1-GFP, respectively. These studies suggest that NM-II isoforms have differential roles in the bleb life cycle. The American Society for Cell Biology 2017-04-15 /pmc/articles/PMC5391180/ /pubmed/28251924 http://dx.doi.org/10.1091/mbc.E16-07-0524 Text en © 2017 Dey, Singh, et al. This article is distributed by The American Society for Cell Biology under license from the author(s). Two months after publication it is available to the public under an Attribution–Noncommercial–Share Alike 3.0 Unported Creative Commons License (http://creativecommons.org/licenses/by-nc-sa/3.0). “ASCB®,” “The American Society for Cell Biology®,” and “Molecular Biology of the Cell®” are registered trademarks of The American Society for Cell Biology. |
spellingShingle | Articles Dey, Sumit K. Singh, Raman K. Chattoraj, Shyamtanu Saha, Shekhar Das, Alakesh Bhattacharyya, Kankan Sengupta, Kaushik Sen, Shamik Jana, Siddhartha S. Differential role of nonmuscle myosin II isoforms during blebbing of MCF-7 cells |
title | Differential role of nonmuscle myosin II isoforms during blebbing of MCF-7 cells |
title_full | Differential role of nonmuscle myosin II isoforms during blebbing of MCF-7 cells |
title_fullStr | Differential role of nonmuscle myosin II isoforms during blebbing of MCF-7 cells |
title_full_unstemmed | Differential role of nonmuscle myosin II isoforms during blebbing of MCF-7 cells |
title_short | Differential role of nonmuscle myosin II isoforms during blebbing of MCF-7 cells |
title_sort | differential role of nonmuscle myosin ii isoforms during blebbing of mcf-7 cells |
topic | Articles |
url | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC5391180/ https://www.ncbi.nlm.nih.gov/pubmed/28251924 http://dx.doi.org/10.1091/mbc.E16-07-0524 |
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