Membrane structure and internalization dynamics of human Flower isoforms hFWE3 and hFWE4 indicate a conserved endocytic role for hFWE4.
Membrane structure and internalization dynamics of human Flower isoforms hFWE3 and hFWE4 indicate a conserved endocytic role for hFWE4.
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HFWE3和hFWE4亚型的膜结构和内化动力学表明hFWE4具有保守的内吞作用。
DOI：
10.1016/j.jbc.2023.104945
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发表时间：
2023-08
影响因子：
4.8
通讯作者：
Hansen, Laura A.
中科院分区：
文献类型：
--
作者：
Rudd, Justin C.;Maity, Sibaprasad;Grunkemeyer, James A.;Snyder, Joshua C.;Lovas, Sandor;Hansen, Laura A.
人花（hFWE）同种型hFWE 1 -4是推定的跨膜（TM）蛋白，据报道其在细胞竞争期间通过其C-末端尾的细胞外展示介导适合度比较。同种型拓扑结构、亚细胞定位和质膜呈递的持续时间对该功能至关重要。然而，关于果蝇和小鼠FWE的结构仍然存在分歧，并且缺乏hFWE同种型亚细胞定位或膜结构的实验证据。在这里，我们使用AlphaFold 2和随后的基于分子动力学的结构预测构建表位标记的hFWE 3和hFWE 4，最丰富的人类同种型，用于实验测定其结构和内化动力学。我们证明，hFWE 3驻留在内质网（ER）的膜，而hFWE 4部分共定位与Rab 4-，Rab 5-，Rab 11-阳性囊泡以及与质膜。一系列成像技术显示，hFWE 4在胞质溶胶内定位N-和C-末端尾和第二和第三TM区段之间的环，而第一和第二以及第三和第四TM区段之间的小（4- 12 aa）环暴露于细胞外空间或胞质囊泡的内腔内。类似地，我们发现hFWE 3在胞质溶胶中定位N-和C-末端尾，而TM结构域之间的短环延伸到ER腔中。最后，我们证明，hFWE 4只存在于细胞表面短暂，并迅速内化的AP-2和动力蛋白-1依赖的方式。总的来说，这些数据与hFWE 4在内吞过程中的保守作用一致。
Human Flower (hFWE) isoforms hFWE1-4 are putative transmembrane (TM) proteins that reportedly mediate fitness comparisons during cell competition through extracellular display of their C-terminal tails. Isoform topology, subcellular localization, and duration of plasma membrane presentation are essential to this function. However, disagreement persists regarding the structure of orthologous fly and mouse FWEs, and experimental evidence for hFWE isoform subcellular localization or membrane structure is lacking. Here, we used AlphaFold2 and subsequent molecular dynamics-based structural predictions to construct epitope-tagged hFWE3 and hFWE4, the most abundant human isoforms, for experimental determination of their structure and internalization dynamics. We demonstrate that hFWE3 resides in the membrane of the endoplasmic reticulum (ER), while hFWE4 partially colocalizes with Rab4-, Rab5-, and Rab11-positive vesicles as well as with the plasma membrane. An array of imaging techniques revealed that hFWE4 positions both N- and C-terminal tails and a loop between second and third TM segments within the cytosol, while small (4–12aa) loops between the first and second and the third and fourth TM segments are either exposed to the extracellular space or within the lumen of cytoplasmic vesicles. Similarly, we found hFWE3 positions both N- and C-terminal tails in the cytosol, while a short loop between TM domains extends into the ER lumen. Finally, we demonstrate that hFWE4 exists only transiently at the cell surface and is rapidly internalized in an AP-2- and dynamin-1-dependent manner. Collectively, these data are consistent with a conserved role for hFWE4 in endocytic processes.
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影响因子：
3
作者：
Krieger E;Vriend G
通讯作者：
Vriend G
长阅读测序揭示了人体组织中的转录组变化。
影响因子：
64.8
作者：
Glinos, Dafni A.;Garborcauskas, Garrett;Hoffman, Paul;Ehsan, Nava;Jiang, Lihua;Gokden, Alper;Dai, Xiaoguang;Aguet, Francois;Brown, Kathleen L.;Garimella, Kiran;Bowers, Tera;Costello, Maura;Ardlie, Kristin;Jian, Ruiqi;Tucker, Nathan R.;Ellinor, Patrick T.;Harrington, Eoghan D.;Tang, Hua;Snyder, Michael;Juul, Sissel;Mohammadi, Pejman;MacArthur, Daniel G.;Lappalainen, Tuuli;Cummings, Beryl
通讯作者：
Cummings, Beryl
影响因子：
64.8
作者：
Jumper J;Evans R;Pritzel A;Green T;Figurnov M;Ronneberger O;Tunyasuvunakool K;Bates R;Žídek A;Potapenko A;Bridgland A;Meyer C;Kohl SAA;Ballard AJ;Cowie A;Romera-Paredes B;Nikolov S;Jain R;Adler J;Back T;Petersen S;Reiman D;Clancy E;Zielinski M;Steinegger M;Pacholska M;Berghammer T;Bodenstein S;Silver D;Vinyals O;Senior AW;Kavukcuoglu K;Kohli P;Hassabis D
通讯作者：
Hassabis D
影响因子：
3.3
作者：
McEwen AE;Maher MT;Mo R;Gottardi CJ
通讯作者：
Gottardi CJ
影响因子：
64.5
作者：
Merino MM;Rhiner C;Lopez-Gay JM;Buechel D;Hauert B;Moreno E
通讯作者：
Moreno E