The carboxy terminal coiled-coil modulates Orai1 internalization during meiosis
| العنوان: | The carboxy terminal coiled-coil modulates Orai1 internalization during meiosis |
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| المؤلفون: | Rawad Hodeify, Maya Dib, Ethel Alcantara-Adap, Raphael Courjaret, Nancy Nader, Cleo Z. Reyes, Ayat S. Hammad, Satanay Hubrack, Fang Yu, Khaled Machaca |
| المصدر: | Scientific Reports Scientific Reports, Vol 11, Iss 1, Pp 1-12 (2021) |
| سنة النشر: | 2020 |
| مصطلحات موضوعية: | Microscopy, Confocal, ORAI1 Protein, Science, education, Caveolin 1, Membrane Proteins, Clathrin, Endocytosis, Article, Meiosis, Xenopus laevis, Calcium channels, Mutation, Medicine, Medical biochemistry and metabolomics, Animals, Calcium, Female, Calcium Signaling, rab5 GTP-Binding Proteins |
| الوصف: | Regulation of Ca2+ signaling is critical for the progression of cell division, especially during meiosis to prepare the egg for fertilization. The primary Ca2+ influx pathway in oocytes is Store-Operated Ca2+ Entry (SOCE). SOCE is tightly regulated during meiosis, including internalization of the SOCE channel, Orai1. Orai1 is a four-pass membrane protein with cytosolic N- and C-termini. Orai1 internalization requires a caveolin binding motif (CBM) in the N-terminus as well as the C-terminal cytosolic domain. However, the molecular determinant for Orai1 endocytosis in the C-terminus are not known. Here we show that the Orai1 C-terminus modulates Orai1 endocytosis during meiosis through a structural motif that is based on the strength of the C-terminal intersubunit coiled coil (CC) domains. Deletion mutants show that a minimal C-terminal sequence after transmembrane domain 4 (residues 260–275) supports Orai1 internalization. We refer to this region as the C-terminus Internalization Handle (CIH). Access to CIH however is dependent on the strength of the intersubunit CC. Mutants that increase the stability of the coiled coil prevent internalization independent of specific mutation. We further used human and Xenopus Orai isoforms with different propensity to form C-terminal CC and show a strong correlation between the strength of the CC and Orai internalization. Furthermore, Orai1 internalization does not depend on clathrin, flotillin or PIP2. Collectively these results argue that Orai1 internalization requires both the N-terminal CBM and C-terminal CIH where access to CIH is controlled by the strength of intersubunit C-terminal CC.Other Information Published in: Scientific Reports License: https://creativecommons.org/licenses/by/4.0See article on publisher's website: http://dx.doi.org/10.1038/s41598-021-82048-z |
| تدمد: | 2045-2322 |
| URL الوصول: | https://explore.openaire.eu/search/publication?articleId=doi_dedup___::130e00dd498706df6eef5609cb65c5d3 https://pubmed.ncbi.nlm.nih.gov/33504898 |
| حقوق: | OPEN |
| رقم الأكسشن: | edsair.doi.dedup.....130e00dd498706df6eef5609cb65c5d3 |
| قاعدة البيانات: | OpenAIRE |
| تدمد: | 20452322 |
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