Article ID | Journal | Published Year | Pages | File Type |
---|---|---|---|---|
10766734 | Biochemical and Biophysical Research Communications | 2008 | 7 Pages |
Abstract
We have demonstrated that T-cell receptor ζ (ζ) mRNA with a 562-bp deleted alternatively spliced 3â²-untranslated region (3â²UTR) observed in T cells of patients with systemic lupus erythematosus (SLE) can lead to a reduction in ζ and TCR/CD3 (J. Immunol., 2003 & 2005). To determine the region in ζ mRNA 3â²UTR for the regulation of ζ, ζ mRNA with 3â²UTR truncations ligated into pDON-AI was used to infect murine T-cell hybridoma MA5.8 cells, which do not contain ζ. As a Western blot analysis demonstrated the importance of the regions from +871 to +950, containing conservative sequence 1 (CS1), and +1070 to +1136, containing CS2, for the production of ζ, we constructed MA5.8 mutants carrying ζ mRNA 3â²UTR with deletions of these regions (ÎCS1 and ÎCS2 mutants). Western blot and FACS analyses showed significant reduction in the cell surface ζ and TCR/CD3 in both these mutants, and IL-2 production was decreased, compared with MA5.8 cells transfected with wild-type ζ mRNA. Furthermore, real-time PCR demonstrated the instability of ζ mRNA with 3â²UTR deletions in these MA5.8 mutants. In conclusion, CS1 and CS2 may be responsible for the regulation of ζ and TCR/CD3 through the stability of ζ mRNA in SLE T cells.
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Authors
Kensei Tsuzaka, Yuka Itami, Chika Kumazawa, Miyuki Suzuki, Yumiko Setoyama, Keiko Yoshimoto, Katsuya Suzuki, Tohru Abe, Tsutomu Takeuchi,