The carboxy terminal coiled-coil modulates Orai1 internalization during meiosis
<p dir="ltr">Regulation of Ca<sup>2+</sup> signaling is critical for the progression of cell division, especially during meiosis to prepare the egg for fertilization. The primary Ca<sup>2+</sup> influx pathway in oocytes is Store-Operated Ca<sup>2+</s...
Saved in:
| Main Author: | |
|---|---|
| Other Authors: | , , , , , , , , |
| Published: |
2021
|
| Subjects: | |
| Tags: |
Add Tag
No Tags, Be the first to tag this record!
|
| _version_ | 1864513554945671168 |
|---|---|
| author | Rawad Hodeify (9592365) |
| author2 | Maya Dib (9592362) Ethel Alcantara-Adap (14152851) Raphael Courjaret (9592368) Nancy Nader (175869) Cleo Z. Reyes (14152854) Ayat S. Hammad (9592374) Satanay Hubrack (194364) Fang Yu (156838) Khaled Machaca (14151234) |
| author2_role | author author author author author author author author author |
| author_facet | Rawad Hodeify (9592365) Maya Dib (9592362) Ethel Alcantara-Adap (14152851) Raphael Courjaret (9592368) Nancy Nader (175869) Cleo Z. Reyes (14152854) Ayat S. Hammad (9592374) Satanay Hubrack (194364) Fang Yu (156838) Khaled Machaca (14151234) |
| author_role | author |
| dc.creator.none.fl_str_mv | Rawad Hodeify (9592365) Maya Dib (9592362) Ethel Alcantara-Adap (14152851) Raphael Courjaret (9592368) Nancy Nader (175869) Cleo Z. Reyes (14152854) Ayat S. Hammad (9592374) Satanay Hubrack (194364) Fang Yu (156838) Khaled Machaca (14151234) |
| dc.date.none.fl_str_mv | 2021-01-27T00:00:00Z |
| dc.identifier.none.fl_str_mv | 10.1038/s41598-021-82048-z |
| dc.relation.none.fl_str_mv | https://figshare.com/articles/journal_contribution/The_carboxy_terminal_coiled-coil_modulates_Orai1_internalization_during_meiosis/21598194 |
| dc.rights.none.fl_str_mv | CC BY 4.0 info:eu-repo/semantics/openAccess |
| dc.subject.none.fl_str_mv | Biomedical and clinical sciences Medical biochemistry and metabolomics Ca2+ Signaling Store-Operated Calcium Entry (SOCE) Orai1 Endocytosis Internalization Handle (CIH) Xenopus |
| dc.title.none.fl_str_mv | The carboxy terminal coiled-coil modulates Orai1 internalization during meiosis |
| dc.type.none.fl_str_mv | Text Journal contribution info:eu-repo/semantics/publishedVersion text contribution to journal |
| description | <p dir="ltr">Regulation of Ca<sup>2+</sup> signaling is critical for the progression of cell division, especially during meiosis to prepare the egg for fertilization. The primary Ca<sup>2+</sup> influx pathway in oocytes is Store-Operated Ca<sup>2+</sup> 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 <i>Xenopus</i> 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.</p><h2>Other Information</h2><p dir="ltr">Published in: Scientific Reports<br>License: <a href="https://creativecommons.org/licenses/by/4.0" target="_blank">https://creativecommons.org/licenses/by/4.0</a><br>See article on publisher's website: <a href="http://dx.doi.org/10.1038/s41598-021-82048-z" target="_blank">http://dx.doi.org/10.1038/s41598-021-82048-z</a></p> |
| eu_rights_str_mv | openAccess |
| id | Manara2_5bfb8c9431c9ae448ddf78b73403ee7b |
| identifier_str_mv | 10.1038/s41598-021-82048-z |
| network_acronym_str | Manara2 |
| network_name_str | Manara2 |
| oai_identifier_str | oai:figshare.com:article/21598194 |
| publishDate | 2021 |
| repository.mail.fl_str_mv | |
| repository.name.fl_str_mv | |
| repository_id_str | |
| rights_invalid_str_mv | CC BY 4.0 |
| spelling | The carboxy terminal coiled-coil modulates Orai1 internalization during meiosisRawad Hodeify (9592365)Maya Dib (9592362)Ethel Alcantara-Adap (14152851)Raphael Courjaret (9592368)Nancy Nader (175869)Cleo Z. Reyes (14152854)Ayat S. Hammad (9592374)Satanay Hubrack (194364)Fang Yu (156838)Khaled Machaca (14151234)Biomedical and clinical sciencesMedical biochemistry and metabolomicsCa2+Signaling Store-Operated Calcium Entry (SOCE)Orai1EndocytosisInternalization Handle (CIH)Xenopus<p dir="ltr">Regulation of Ca<sup>2+</sup> signaling is critical for the progression of cell division, especially during meiosis to prepare the egg for fertilization. The primary Ca<sup>2+</sup> influx pathway in oocytes is Store-Operated Ca<sup>2+</sup> 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 <i>Xenopus</i> 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.</p><h2>Other Information</h2><p dir="ltr">Published in: Scientific Reports<br>License: <a href="https://creativecommons.org/licenses/by/4.0" target="_blank">https://creativecommons.org/licenses/by/4.0</a><br>See article on publisher's website: <a href="http://dx.doi.org/10.1038/s41598-021-82048-z" target="_blank">http://dx.doi.org/10.1038/s41598-021-82048-z</a></p>2021-01-27T00:00:00ZTextJournal contributioninfo:eu-repo/semantics/publishedVersiontextcontribution to journal10.1038/s41598-021-82048-zhttps://figshare.com/articles/journal_contribution/The_carboxy_terminal_coiled-coil_modulates_Orai1_internalization_during_meiosis/21598194CC BY 4.0info:eu-repo/semantics/openAccessoai:figshare.com:article/215981942021-01-27T00:00:00Z |
| spellingShingle | The carboxy terminal coiled-coil modulates Orai1 internalization during meiosis Rawad Hodeify (9592365) Biomedical and clinical sciences Medical biochemistry and metabolomics Ca2+ Signaling Store-Operated Calcium Entry (SOCE) Orai1 Endocytosis Internalization Handle (CIH) Xenopus |
| status_str | publishedVersion |
| title | 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_short | The carboxy terminal coiled-coil modulates Orai1 internalization during meiosis |
| title_sort | The carboxy terminal coiled-coil modulates Orai1 internalization during meiosis |
| topic | Biomedical and clinical sciences Medical biochemistry and metabolomics Ca2+ Signaling Store-Operated Calcium Entry (SOCE) Orai1 Endocytosis Internalization Handle (CIH) Xenopus |