Asymmetric tethering by exocyst in vitro requires a Rab GTPase, a v-SNARE and a Sac1-sensitive phosphoinositide lipid
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Abstract
Tethering factors play a critical role in deciphering the correct combination of vesicle and target membrane for subsequent fusion. The exocyst plays a central role in tethering post-Golgi vesicles to the plasma membrane, although the mechanism by which this occurs is poorly understood. We recently established an assay for measuring exocyst-mediated vesicle tethering in vitro and we have adapted this assay to examine the ability of exocyst to tether vesicles in an asymmetric fashion. We demonstrate that exocyst differs from another post-Golgi vesicle tethering protein, Sro7, in that it is fully capable of tethering vesicles with functional Rab GTPase, Sec4, to vesicles lacking a functional Rab GTPase. Using this assay, we show that exocyst requires both the Rab and R-SNARE, Snc1, to be present on the same membrane surface. In contrast, using Sac1 phosphatase treatment, we demonstrate a likely role for phosphoinositides on the opposing Rab-deficient membrane. This suggests a specific model for exocyst orientation and its points of contact between membranes during heterotypic tethering of post-Golgi vesicles with the plasma membrane.
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This Zenodo record is a permanently preserved version of a Structured PREreview. You can view the complete PREreview at https://prereview.org/reviews/10093809.
Does the introduction explain the objective of the research presented in the preprint? YesAre the methods well-suited for this research? Highly appropriateAre the conclusions supported by the data? Somewhat supported It is generalized that the tethering reaction to the plasma membrane need both Rabs and R-SNAREs, while it was study only on representative per each of them such as …This Zenodo record is a permanently preserved version of a Structured PREreview. You can view the complete PREreview at https://prereview.org/reviews/10093809.
Does the introduction explain the objective of the research presented in the preprint? YesAre the methods well-suited for this research? Highly appropriateAre the conclusions supported by the data? Somewhat supported It is generalized that the tethering reaction to the plasma membrane need both Rabs and R-SNAREs, while it was study only on representative per each of them such as Sec4 and Sso1.Are the data presentations, including visualizations, well-suited to represent the data? Somewhat appropriate and clear The images from the microscopy need to have higher quality and scale bars. Figure legends are lacking the number of samples analyzed.How clearly do the authors discuss, explain, and interpret their findings and potential next steps for the research? Neither clearly nor unclearly They need to discuss more the results based on similar findings in other model organisms.Is the preprint likely to advance academic knowledge? Moderately likelyWould it benefit from language editing? Yes It might need more logical flow and development of the paragraph as sometimes the sentences are short.Would you recommend this preprint to others? Yes, but it needs to be improvedIs it ready for attention from an editor, publisher or broader audience? Yes, after minor changes It might need work regarding the discussion of the results.Competing interests
The author declares that they have no competing interests.
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