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探究将外排 Rab 重定向到内吞小泡的后果。

Exploring the consequences of redirecting an exocytic Rab onto endocytic vesicles.

机构信息

Department of Cellular and Molecular Medicine, University of California, San Diego, La Jolla, CA 92093-0644.

Nikon Imaging Center, University of California, San Diego, La Jolla, CA 92093-0694.

出版信息

Mol Biol Cell. 2023 May 1;34(5):ar38. doi: 10.1091/mbc.E23-01-0037. Epub 2023 Mar 1.

DOI:10.1091/mbc.E23-01-0037
PMID:36857153
原文链接:https://pmc.ncbi.nlm.nih.gov/articles/PMC10162416/
Abstract

Bidirectional vesicular traffic links compartments along the exocytic and endocytic pathways. Rab GTPases have been implicated in specifying the direction of vesicular transport. To explore this possibility, we sought to redirect an exocytic Rab, Sec4, onto endocytic vesicles by fusing the catalytic domain of the Sec4 GEF, Sec2, onto the CUE localization domain of Vps9, a GEF for the endocytic Rab Ypt51. The Sec2GEF-GFP-CUE construct localized to bright puncta predominantly near sites of polarized growth, and this localization was dependent on the ability of the CUE domain to bind to the ubiquitin moieties added to the cytoplasmic tails of proteins destined for endocytic internalization. Sec4 and Sec4 effectors were recruited to these puncta with various efficiencies. Cells expressing Sec2GEF-GFP-CUE grew surprisingly well and secreted protein at near-normal efficiency, implying that Golgi-derived secretory vesicles were delivered to polarized sites of cell growth despite the misdirection of Sec4 and its effectors. A low efficiency mechanism for localization of Sec2 to secretory vesicles that is independent of known cues might be responsible. In total, the results suggest that while Rabs may play a critical role in specifying the direction of vesicular transport, cells are remarkably tolerant of Rab misdirection.

摘要

双向囊泡运输将沿着胞吐和胞吞途径的隔室连接起来。Rab GTPases 被认为参与了囊泡运输方向的指定。为了探索这种可能性，我们试图通过将 Sec4 GEF Sec2 的催化结构域融合到 Vps9 的 CUE 定位结构域上，将外排 Rab Sec4 重新导向内吞囊泡，Vps9 是内吞 Rab Ypt51 的 GEF。Sec2GEF-GFP-CUE 结构定位到明亮的斑点，主要靠近极化生长的部位，这种定位依赖于 CUE 结构域结合到细胞质尾部添加的泛素部分的能力，这些尾部蛋白是内吞内化的。Sec4 和 Sec4 效应物以不同的效率被招募到这些斑点。表达 Sec2GEF-GFP-CUE 的细胞生长得非常好，并且以接近正常的效率分泌蛋白质，这意味着尽管 Sec4 和其效应物被误导，但高尔基体衍生的分泌囊泡被递送到极化的细胞生长部位。可能是一种独立于已知线索的 Sec2 到分泌囊泡的低效率定位机制负责。总的来说，这些结果表明，虽然 Rab 可能在指定囊泡运输方向方面发挥关键作用，但细胞对 Rab 错误定位具有很强的耐受性。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/f18a/10162416/2b45b93bc605/mbc-34-ar38-g006.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/f18a/10162416/d04a047e13f0/mbc-34-ar38-g001.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/f18a/10162416/26d9dccbd7cd/mbc-34-ar38-g002.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/f18a/10162416/37c930aba239/mbc-34-ar38-g003.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/f18a/10162416/3fcdd05a382e/mbc-34-ar38-g004.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/f18a/10162416/08e3ec44224a/mbc-34-ar38-g005.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/f18a/10162416/2b45b93bc605/mbc-34-ar38-g006.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/f18a/10162416/d04a047e13f0/mbc-34-ar38-g001.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/f18a/10162416/26d9dccbd7cd/mbc-34-ar38-g002.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/f18a/10162416/37c930aba239/mbc-34-ar38-g003.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/f18a/10162416/3fcdd05a382e/mbc-34-ar38-g004.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/f18a/10162416/08e3ec44224a/mbc-34-ar38-g005.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/f18a/10162416/2b45b93bc605/mbc-34-ar38-g006.jpg

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