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ID 114181
Author
Utami, Trianna W. The University of Tokushima
Hagita, Hiroko The University of Tokushima
Yanuaryska, Ryna D. The University of Tokushima
Keywords
DNA methylation
epigenetic
regulation
gain-of-function
histone modification
Sp6
Content Type
Journal Article
Description
To investigate the function of specificity protein 6 (SP6) transcription factor by gain-of-function procedure, we established cytomegalovirus (CMV) promoter-driven Sp6 stable transformants, C9 cells, using dental epithelial-derived cells. Initially, C9 cells produced a significant amount of SP6 protein. However, SP6 expression was reduced in these cells upon long-term culture. We could detect Sp6 transcripts in C9 cells by RT-PCR throughout the passages, although the CMV promoter is known to be epigenetically silenced. We recently found that SP6 was a short-lived protein that was degraded by a ubiquitin-independent proteasome pathway, although it is yet unclear how Sp6 expression was regulated during culture. Thus, we studied the possibility of epigenetic regulation of Sp6 expression. Comparative analysis of endogenous and exogenous Sp6 mRNA expressions demonstrated the specific down-regulation of exogenous Sp6 mRNA levels during culture passages. A DNA methyltransferase inhibitor, 5-Aza-2'-deoxycytidine (5AC), and a histone deacetylase inhibitor, valproic acid (VPA), enhanced or induced SP6 protein expression up to passage 28 without enhancing the mRNA level. The dramatic up-regulation of exogenous Sp6 mRNA was uniquely observed only at passage 50 by 5AC or VPA treatment. These findings indicate that multiple epigenetic regulatory mechanisms operate to fine-tune Sp6 expression during long-term culture.
Journal Title
The Indonesian Journal of Dental Research
ISSN
20878710
24423300
Publisher
Universitas Gadjah Mada
Volume
1
Issue
3
Start Page
134
End Page
142
Published Date
2011
Rights
This work is licensed under a Creative Commons Attribution-ShareAlike 4.0 International License(https://creativecommons.org/licenses/by-sa/4.0/).
EDB ID
DOI (Published Version)
URL ( Publisher's Version )
FullText File
language
eng
TextVersion
Publisher
departments
Oral Sciences