Abstract
We have demonstrated that T-cell receptor ζ (ζ) mRNA with a 562-bp deleted alternatively spliced 3′-untranslated region (3′UTR) 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 3′UTR for the regulation of ζ, ζ mRNA with 3′UTR 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 3′UTR with deletions of these regions (ΔCS1 and ΔCS2 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 3′UTR 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.
| Original language | English |
|---|---|
| Pages (from-to) | 311-317 |
| Number of pages | 7 |
| Journal | Biochemical and Biophysical Research Communications |
| Volume | 367 |
| Issue number | 2 |
| DOIs | |
| Publication status | Published - 2008 Mar 7 |
| Externally published | Yes |
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Keywords
- 3′UTR
- Autoimmune disease
- Conservative sequence
- IL-2
- MA5.8 cells
- mRNA stability
- Signal transduction
- Systemic lupus erythematosus
- T-cell receptor
- TCRζ
ASJC Scopus subject areas
- Biochemistry
- Biophysics
- Molecular Biology
Cite this
Conservative sequences in 3′UTR of TCRζ mRNA regulate TCRζ in SLE T cells. / Tsuzaka, Kensei; Itami, Yuka; Kumazawa, Chika; Suzuki, Miyuki; Setoyama, Yumiko; Yoshimoto, Keiko; Suzuki, Katsuya; Abe, Tohru; Takeuchi, Tsutomu.
In: Biochemical and Biophysical Research Communications, Vol. 367, No. 2, 07.03.2008, p. 311-317.Research output: Contribution to journal › Article
}
TY - JOUR
T1 - Conservative sequences in 3′UTR of TCRζ mRNA regulate TCRζ in SLE T cells
AU - Tsuzaka, Kensei
AU - Itami, Yuka
AU - Kumazawa, Chika
AU - Suzuki, Miyuki
AU - Setoyama, Yumiko
AU - Yoshimoto, Keiko
AU - Suzuki, Katsuya
AU - Abe, Tohru
AU - Takeuchi, Tsutomu
PY - 2008/3/7
Y1 - 2008/3/7
N2 - We have demonstrated that T-cell receptor ζ (ζ) mRNA with a 562-bp deleted alternatively spliced 3′-untranslated region (3′UTR) 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 3′UTR for the regulation of ζ, ζ mRNA with 3′UTR 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 3′UTR with deletions of these regions (ΔCS1 and ΔCS2 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 3′UTR 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.
AB - We have demonstrated that T-cell receptor ζ (ζ) mRNA with a 562-bp deleted alternatively spliced 3′-untranslated region (3′UTR) 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 3′UTR for the regulation of ζ, ζ mRNA with 3′UTR 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 3′UTR with deletions of these regions (ΔCS1 and ΔCS2 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 3′UTR 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.
KW - 3′UTR
KW - Autoimmune disease
KW - Conservative sequence
KW - IL-2
KW - MA5.8 cells
KW - mRNA stability
KW - Signal transduction
KW - Systemic lupus erythematosus
KW - T-cell receptor
KW - TCRζ
UR - http://www.scopus.com/inward/record.url?scp=38349053810&partnerID=8YFLogxK
UR - http://www.scopus.com/inward/citedby.url?scp=38349053810&partnerID=8YFLogxK
U2 - 10.1016/j.bbrc.2007.12.145
DO - 10.1016/j.bbrc.2007.12.145
M3 - Article
VL - 367
SP - 311
EP - 317
JO - Biochemical and Biophysical Research Communications
JF - Biochemical and Biophysical Research Communications
SN - 0006-291X
IS - 2
ER -