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...
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| Format: | Article |
| Language: | English |
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Public Library of Science (PLoS)
2013-01-01
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| Series: | PLoS ONE |
| Online Access: | http://europepmc.org/articles/PMC3776769?pdf=render |
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| author | Takashi Tsukamoto Xianglan Li Hiromi Morita Takashi Minowa Tomoyasu Aizawa Nobutaka Hanagata Makoto Demura |
| author_facet | Takashi Tsukamoto Xianglan Li Hiromi Morita Takashi Minowa Tomoyasu Aizawa Nobutaka Hanagata Makoto Demura |
| author_sort | Takashi Tsukamoto |
| collection | DOAJ |
| 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. |
| first_indexed | 2024-12-13T15:17:10Z |
| format | Article |
| id | doaj.art-27a5571d1d4c4d2a8e015060e46f7cab |
| institution | Directory Open Access Journal |
| issn | 1932-6203 |
| language | English |
| last_indexed | 2024-12-13T15:17:10Z |
| publishDate | 2013-01-01 |
| publisher | Public Library of Science (PLoS) |
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| series | PLoS ONE |
| spelling | doaj.art-27a5571d1d4c4d2a8e015060e46f7cab2022-12-21T23:40:41ZengPublic Library of Science (PLoS)PLoS ONE1932-62032013-01-0189e7583110.1371/journal.pone.0075831Role of S-palmitoylation on IFITM5 for the interaction with FKBP11 in osteoblast cells.Takashi TsukamotoXianglan LiHiromi MoritaTakashi MinowaTomoyasu AizawaNobutaka HanagataMakoto DemuraRecently, 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.http://europepmc.org/articles/PMC3776769?pdf=render |
| spellingShingle | Takashi Tsukamoto Xianglan Li Hiromi Morita Takashi Minowa Tomoyasu Aizawa Nobutaka Hanagata Makoto Demura Role of S-palmitoylation on IFITM5 for the interaction with FKBP11 in osteoblast cells. PLoS ONE |
| 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 |
| url | http://europepmc.org/articles/PMC3776769?pdf=render |
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