RRC ID 44661
Author Tsukamoto T, Li X, Morita H, Minowa T, Aizawa T, Hanagata N, Demura M.
Title Role of S-palmitoylation on IFITM5 for the interaction with FKBP11 in osteoblast cells.
Journal PLoS One
Abstract 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.
Volume 8(9)
Pages e75831
Published 2013
DOI 10.1371/journal.pone.0075831
PII PONE-D-13-16644
PMID 24058703
PMC PMC3776769
MeSH Animals Enzyme Inhibitors / pharmacology Lipoylation / drug effects Lipoylation / physiology* Membrane Proteins / genetics Membrane Proteins / metabolism* Mice Multiprotein Complexes / genetics Multiprotein Complexes / metabolism* Osteoblasts / cytology Osteoblasts / metabolism* Osteogenesis / drug effects Osteogenesis / physiology* Protein Structure, Secondary Protein Structure, Tertiary Tacrolimus Binding Proteins / genetics Tacrolimus Binding Proteins / metabolism*
IF 2.776
Times Cited 18
WOS Category BIOCHEMISTRY & MOLECULAR BIOLOGY
Resource
Human and Animal Cells