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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...

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Autores principales: Dey, Sumit K., Singh, Raman K., Chattoraj, Shyamtanu, Saha, Shekhar, Das, Alakesh, Bhattacharyya, Kankan, Sengupta, Kaushik, Sen, Shamik, Jana, Siddhartha S.
Formato: Online Artículo Texto
Lenguaje:English
Publicado: The American Society for Cell Biology 2017
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.
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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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