Cigarette smoke impairs the endocytotic process in Saccharomyces cerevisiae
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Abstract
The accumulation of misfolded proteins inside the cells has been considered to be an important contributor to the development of cigarette smoke-mediated diseases. Since endocytosis plays a crucial role in protein trafficking and clearance, impaired endocytosis may contribute to cigarette smoke-mediated protein accumulation. Therefore, the current study investigated the effects of cigarette smoke extract (CSE) on the endocytosis process in yeast Saccharomyces cerevisiae . The current study showed that treatment of cells with CSE caused reduced uptake of FM4-64 stain, indicating impaired endocytosis. Further analysis revealed that CSE treatment resulted in a defect in the recruitment of proteins involved in endocytosis. Also, aberrant actin morphology was found upon CSE treatment, which might interfere with vesicle budding from the membrane. Moreover, the current study showed that the PI4,5P2 level in the plasma membrane in CSE treated cells is reduced due to the failed translocation of MSS4 kinase to the membrane. This reduced PI4,5P2 results in aberrant actin morphology. Thus, the current study demonstrates that CSE treatment causes endocytosis defects and provides insight into this defective endocytosis.
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a substantial amount of signal
A quantification would be helpful
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, MSS4-GFP failed to get recruited at the membrane resulting in reduced PI4,5P2 level at the membrane
I see no meaningful difference between the CSE-treated and untreated cells for MSS4-GFP. How are you determining this difference? Is there a quantification?
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exhibited significant diffused cytosolic signals
There does not appear to be a methods section. How were the images taken? For example, was this a slice of a confocal z-stack such that signal in the middle of the cell really is in the cytosol or a wide field image such that the "cytosolic" signal may just be signal from accumulation of Ede1 at the cell surface (aka endocytosis is stuck at Ede1 recruitment step so a lot more enrichment is visible at the top and bottom of the cell). In other words have these images been taken in a way that can distinguish these possibilities?
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different lengths of time
I might be missing it somewhere but the two time points do not appear to be in the figure or figure legend -- from the text it appears maybe the bottom row is 2 hours?
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