Journal
PLOS ONE
Volume 10, Issue 1, Pages -Publisher
PUBLIC LIBRARY SCIENCE
DOI: 10.1371/journal.pone.0116579
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Funding
- Tokyo Biochemical Research Foundation
- Nakatani Foundation for Advancement of Measuring Technologies in Biomedical Engineering
- Asahi Glass Foundation
- Uehara Memorial Foundation
- Takeda Science Foundation
- Kurata Memorial Hitachi Science and Technology Foundation
- Adaptable & Seamless Technology Transfer Program through Target-driven R&D (A-STEP) by the Japan Science and Technology Agency (JST) [AS242Z00789Q]
- Ministry of Education, Culture, Sports, Science and Technology of Japan [23118516, 23114707, 25118512]
- Program for Combined Research Fields from Immunology Frontier Research Center, Osaka University
- [22710185]
- [25830131]
- Grants-in-Aid for Scientific Research [25118512] Funding Source: KAKEN
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Accumulating evidence suggests that Pax5 plays essential roles in B cell lineage commitment. However, molecular mechanisms of B cell-specific expression of Pax5 are not fully understood. Here, we applied insertional chromatin immunoprecipitation iChIP) combined with stable isotope labeling using amino acids in cell culture (SILAC) (iChIP-SILAC) to direct identification of proteins interacting with the promoter region of the endogenous single-copy chicken Pax5 gene. By comparing B cells with macrophage-like cells trans-differentiated by ectopic expression of C/EBP beta, iChIP-SILAC detected B cell-specific interaction of a nuclear protein, Thy28/Thyn1, with the Pax5 1A promoter. Trans-differentiation of B cells into macrophage- like cells caused down-regulation of Thy28 expression. Loss-of-function of Thy28 induced decrease in Pax5 expression and recruitment of myosin-9 (MYH9), one of Thy28-interacting proteins, to the Pax5 1A promoter. Loss-of-function of MYH9 also induced decrease in Pax5 expression. Thus, our analysis revealed that Thy28 is functionally required for B cell-specific expression of Pax5 via recruitment of MYH9 to the Pax5 locus in chicken B cells.
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