Cargando…

Role of S-Palmitoylation on IFITM5 for the Interaction with FKBP11 in Osteoblast Cells

Recently, one of the interferon-induced transmembrane (IFITM) family proteins, IFITM3, has become an important target for the activity against influenza A (H1N1) virus infection. In this protein, a post-translational modification by fatty acids covalently attached to cysteine, termed S-palmitoylatio...

Descripción completa

Detalles Bibliográficos
Autores principales: Tsukamoto, Takashi, Li, Xianglan, Morita, Hiromi, Minowa, Takashi, Aizawa, Tomoyasu, Hanagata, Nobutaka, Demura, Makoto
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Public Library of Science 2013
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC3776769/
https://www.ncbi.nlm.nih.gov/pubmed/24058703
http://dx.doi.org/10.1371/journal.pone.0075831
_version_ 1782284879516401664
author Tsukamoto, Takashi
Li, Xianglan
Morita, Hiromi
Minowa, Takashi
Aizawa, Tomoyasu
Hanagata, Nobutaka
Demura, Makoto
author_facet Tsukamoto, Takashi
Li, Xianglan
Morita, Hiromi
Minowa, Takashi
Aizawa, Tomoyasu
Hanagata, Nobutaka
Demura, Makoto
author_sort Tsukamoto, Takashi
collection PubMed
description Recently, one of the interferon-induced transmembrane (IFITM) family proteins, IFITM3, has become an important target for the activity against influenza A (H1N1) virus infection. In this protein, a post-translational modification by fatty acids covalently attached to cysteine, termed S-palmitoylation, plays a crucial role for the antiviral activity. IFITM3 possesses three cysteine residues for the S-palmitoylation in the first transmembrane (TM1) domain and in the cytoplasmic (CP) loop. Because these cysteines are well conserved in the mammalian IFITM family proteins, the S-palmitoylation on these cysteines is significant for their functions. IFITM5 is another IFITM family protein and interacts with the FK506-binding protein 11 (FKBP11) to form a higher-order complex in osteoblast cells, which induces the expression of immunologically relevant genes. In this study, we investigated the role played by S-palmitoylation of IFITM5 in its interaction with FKBP11 in the cells, because this interaction is a key process for the gene expression. Our investigations using an established reporter, 17-octadecynoic acid (17-ODYA), and an inhibitor for the S-palmitoylation, 2-bromopalmitic acid (2BP), revealed that IFITM5 was S-palmitoylated in addition to IFITM3. Specifically, we found that cysteine residues in the TM1 domain and in the CP loop were S-palmitoylated in IFITM5. Then, we revealed by immunoprecipitation and western blot analyses that the interaction of IFITM5 with FKBP11 was inhibited in the presence of 2BP. The mutant lacking the S-palmitoylation site in the TM1 domain lost the interaction with FKBP11. These results indicate that the S-palmitoylation on IFITM5 promotes the interaction with FKBP11. Finally, we investigated bone nodule formation in osteoblast cells in the presence of 2BP, because IFITM5 was originally identified as a bone formation factor. The experiment resulted in a morphological aberration of the bone nodule. This also indicated that the S-palmitoylation contributes to bone formation.
format Online
Article
Text
id pubmed-3776769
institution National Center for Biotechnology Information
language English
publishDate 2013
publisher Public Library of Science
record_format MEDLINE/PubMed
spelling pubmed-37767692013-09-20 Role of S-Palmitoylation on IFITM5 for the Interaction with FKBP11 in Osteoblast Cells Tsukamoto, Takashi Li, Xianglan Morita, Hiromi Minowa, Takashi Aizawa, Tomoyasu Hanagata, Nobutaka Demura, Makoto PLoS One Research Article Recently, one of the interferon-induced transmembrane (IFITM) family proteins, IFITM3, has become an important target for the activity against influenza A (H1N1) virus infection. In this protein, a post-translational modification by fatty acids covalently attached to cysteine, termed S-palmitoylation, plays a crucial role for the antiviral activity. IFITM3 possesses three cysteine residues for the S-palmitoylation in the first transmembrane (TM1) domain and in the cytoplasmic (CP) loop. Because these cysteines are well conserved in the mammalian IFITM family proteins, the S-palmitoylation on these cysteines is significant for their functions. IFITM5 is another IFITM family protein and interacts with the FK506-binding protein 11 (FKBP11) to form a higher-order complex in osteoblast cells, which induces the expression of immunologically relevant genes. In this study, we investigated the role played by S-palmitoylation of IFITM5 in its interaction with FKBP11 in the cells, because this interaction is a key process for the gene expression. Our investigations using an established reporter, 17-octadecynoic acid (17-ODYA), and an inhibitor for the S-palmitoylation, 2-bromopalmitic acid (2BP), revealed that IFITM5 was S-palmitoylated in addition to IFITM3. Specifically, we found that cysteine residues in the TM1 domain and in the CP loop were S-palmitoylated in IFITM5. Then, we revealed by immunoprecipitation and western blot analyses that the interaction of IFITM5 with FKBP11 was inhibited in the presence of 2BP. The mutant lacking the S-palmitoylation site in the TM1 domain lost the interaction with FKBP11. These results indicate that the S-palmitoylation on IFITM5 promotes the interaction with FKBP11. Finally, we investigated bone nodule formation in osteoblast cells in the presence of 2BP, because IFITM5 was originally identified as a bone formation factor. The experiment resulted in a morphological aberration of the bone nodule. This also indicated that the S-palmitoylation contributes to bone formation. Public Library of Science 2013-09-18 /pmc/articles/PMC3776769/ /pubmed/24058703 http://dx.doi.org/10.1371/journal.pone.0075831 Text en © 2013 Tsukamoto et al http://creativecommons.org/licenses/by/4.0/ This is an open-access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are properly credited.
spellingShingle Research Article
Tsukamoto, Takashi
Li, Xianglan
Morita, Hiromi
Minowa, Takashi
Aizawa, Tomoyasu
Hanagata, Nobutaka
Demura, Makoto
Role of S-Palmitoylation on IFITM5 for the Interaction with FKBP11 in Osteoblast Cells
title Role of S-Palmitoylation on IFITM5 for the Interaction with FKBP11 in Osteoblast Cells
title_full Role of S-Palmitoylation on IFITM5 for the Interaction with FKBP11 in Osteoblast Cells
title_fullStr Role of S-Palmitoylation on IFITM5 for the Interaction with FKBP11 in Osteoblast Cells
title_full_unstemmed Role of S-Palmitoylation on IFITM5 for the Interaction with FKBP11 in Osteoblast Cells
title_short Role of S-Palmitoylation on IFITM5 for the Interaction with FKBP11 in Osteoblast Cells
title_sort role of s-palmitoylation on ifitm5 for the interaction with fkbp11 in osteoblast cells
topic Research Article
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC3776769/
https://www.ncbi.nlm.nih.gov/pubmed/24058703
http://dx.doi.org/10.1371/journal.pone.0075831
work_keys_str_mv AT tsukamototakashi roleofspalmitoylationonifitm5fortheinteractionwithfkbp11inosteoblastcells
AT lixianglan roleofspalmitoylationonifitm5fortheinteractionwithfkbp11inosteoblastcells
AT moritahiromi roleofspalmitoylationonifitm5fortheinteractionwithfkbp11inosteoblastcells
AT minowatakashi roleofspalmitoylationonifitm5fortheinteractionwithfkbp11inosteoblastcells
AT aizawatomoyasu roleofspalmitoylationonifitm5fortheinteractionwithfkbp11inosteoblastcells
AT hanagatanobutaka roleofspalmitoylationonifitm5fortheinteractionwithfkbp11inosteoblastcells
AT demuramakoto roleofspalmitoylationonifitm5fortheinteractionwithfkbp11inosteoblastcells