• Title/Summary/Keyword: Fusion-splicing

Search Result 22, Processing Time 0.018 seconds

Identification and Characterization of Alternative Promoters of the Rice MAP Kinase Gene OsBWMK1

  • Koo, Sung Cheol;Choi, Man Soo;Chun, Hyun Jin;Park, Hyeong Cheol;Kang, Chang Ho;Shim, Sang In;Chung, Jong Il;Cheong, Yong Hwa;Lee, Sang Yeol;Yun, Dae-Jin;Chung, Woo Sik;Cho, Moo Je;Kim, Min Chul
    • Molecules and Cells
    • /
    • v.27 no.4
    • /
    • pp.467-473
    • /
    • 2009
  • Our previous study suggested that OsBWMK1, a gene which encodes a member of the rice MAP kinase family, generates transcript variants which show distinct expression patterns in response to environmental stresses. The transcript variants are generated by alternative splicing and by use of alternative promoters. To test whether the two alternative promoters, pOsBWMK1L (promoter for the OsBWMK1L splice variant) and pOsBWMK1S (promoter for the OsBWMK1S splice variant), are biologically functional, we analyzed transgenic plants expressing GUS fusion constructs for each promoter. Both pOsBWMK1L and pOsBWMK1S are biologically active, although the activity of pOsBWMK1S is lower than that of pOsBWMK1L. Histochemical analysis revealed that pOsBWMK1L is constitutively active in most tissues at various developmental stages in rice and Arabidopsis, whereas pOsBWMK1S activity is spatially and temporally restricted. Furthermore, the expression of pOsBWMK1S::GUS was upregulated in response to hydrogen peroxide, a plant defense signaling molecule, in both plant species. These results suggest that the differential expression of OsBWMK1 splice variants is the result of alternative promoter usage and, moreover, that the mechanisms controlling OsBWMK1 gene expression are conserved in both monocot and dicot plants.

Up-regulation of Pluripotency-related Genes in Human Amniotic Fluid-derived Stem Cells by ESRRB Conjugated with Cell-Penetrating Peptide (인간 양수 유래 줄기세포에서 세포투과단백질을 이용한 ESRRB 단백질의 직접도입에 의한 전분화능 관련 유전자의 발현 조절)

  • Jo, Jung-Hyun;Lee, Yu-Sun;Oh, Mi-Hee;Ko, Jung-Jae;Cheon, Yong-Pil;Lee, Dong-Ryul
    • Development and Reproduction
    • /
    • v.14 no.4
    • /
    • pp.243-251
    • /
    • 2010
  • ESRRB (Estrogen related receptor $\beta$) is an orphan receptor, and have a role on maintaining the undifferentiated state and self-renewal of pluripotent stem cell as a transcription factor which regulates the expression of OCT4 and NANOG genes. Also, Feng et al. (2009) reported that Esrrb, Oct4 and Sox2 could induce pluripotent stem cell from somatic cells. The aim of the present study was to develop the direct delivery system of human ESRRB protein into human amniotic fluid-derived stem cells (AFSCs) and to analyze the effect of ESRRB on the regulation of pluripotency-related genes. Human ESRRB has three isoforms arisen by alternative splicing. We cloned short-form ESRRB and made a fusion protein of ESRRB and R7 for an efficient protein transfer to cell. R7 as cell-penetrating peptide(CPP) can help to transfer ESRRB into cells. R7-ESRRB-His6 protein was observed in the cytoplasm and nuclei within 5 hours after treatment. Also, we could observe R7-ESRRB-His6 protein only in the nuclei within 24 hours. Realtime PCR showed that ESRRB increased expression of OCT4 and NANOG as well as SOX2 gene. Therefore, we demonstrated that R7-ESRRB-His6 proteins were efficiently transferred into the nuclei of AFSCs and work well as a possible transcription factor.