The carboxy terminal coiled-coil modulates Orai1 internalization during meiosis

Abstract 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...

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Autores principales:	Rawad Hodeify, Maya Dib, Ethel Alcantara-Adap, Raphael Courjaret, Nancy Nader, Cleo Z. Reyes, Ayat S. Hammad, Satanay Hubrack, Fang Yu, Khaled Machaca
Formato:	article
Lenguaje:	EN
Publicado:	Nature Portfolio 2021
Materias:	Medicine R Science Q
Acceso en línea:	https://doaj.org/article/8748cd8b3cdf4c24a5978c063351d052
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spelling	oai:doaj.org-article:8748cd8b3cdf4c24a5978c063351d0522021-12-02T14:16:49ZThe carboxy terminal coiled-coil modulates Orai1 internalization during meiosis10.1038/s41598-021-82048-z2045-2322https://doaj.org/article/8748cd8b3cdf4c24a5978c063351d0522021-01-01T00:00:00Zhttps://doi.org/10.1038/s41598-021-82048-zhttps://doaj.org/toc/2045-2322Abstract 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.Rawad HodeifyMaya DibEthel Alcantara-AdapRaphael CourjaretNancy NaderCleo Z. ReyesAyat S. HammadSatanay HubrackFang YuKhaled MachacaNature PortfolioarticleMedicineRScienceQENScientific Reports, Vol 11, Iss 1, Pp 1-12 (2021)
institution	DOAJ
collection	DOAJ
language	EN
topic	Medicine R Science Q
spellingShingle	Medicine R Science Q Rawad Hodeify Maya Dib Ethel Alcantara-Adap Raphael Courjaret Nancy Nader Cleo Z. Reyes Ayat S. Hammad Satanay Hubrack Fang Yu Khaled Machaca The carboxy terminal coiled-coil modulates Orai1 internalization during meiosis
description	Abstract 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.
format	article
author	Rawad Hodeify Maya Dib Ethel Alcantara-Adap Raphael Courjaret Nancy Nader Cleo Z. Reyes Ayat S. Hammad Satanay Hubrack Fang Yu Khaled Machaca
author_facet	Rawad Hodeify Maya Dib Ethel Alcantara-Adap Raphael Courjaret Nancy Nader Cleo Z. Reyes Ayat S. Hammad Satanay Hubrack Fang Yu Khaled Machaca
author_sort	Rawad Hodeify
title	The carboxy terminal coiled-coil modulates Orai1 internalization during meiosis
title_short	The carboxy terminal coiled-coil modulates Orai1 internalization during meiosis
title_full	The carboxy terminal coiled-coil modulates Orai1 internalization during meiosis
title_fullStr	The carboxy terminal coiled-coil modulates Orai1 internalization during meiosis
title_full_unstemmed	The carboxy terminal coiled-coil modulates Orai1 internalization during meiosis
title_sort	carboxy terminal coiled-coil modulates orai1 internalization during meiosis
publisher	Nature Portfolio
publishDate	2021
url	https://doaj.org/article/8748cd8b3cdf4c24a5978c063351d052
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The carboxy terminal coiled-coil modulates Orai1 internalization during meiosis

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