RRC ID |
2433
|
Author |
Kuroda T, Tada M, Kubota H, Kimura H, Hatano SY, Suemori H, Nakatsuji N, Tada T.
|
Title |
Octamer and Sox elements are required for transcriptional cis regulation of Nanog gene expression.
|
Journal |
Mol Cell Biol
|
Abstract |
The pluripotential cell-specific gene Nanog encodes a homeodomain-bearing transcription factor required for maintaining the undifferentiated state of stem cells. However, the molecular mechanisms that regulate Nanog gene expression are largely unknown. To address this important issue, we used luciferase assays to monitor the relative activities of deletion fragments from the 5'-flanking region of the gene. An adjacent pair of highly conserved Octamer- and Sox-binding sites was found to be essential for activating pluripotential state-specific gene expression. Furthermore, the 5'-end fragment encompassing the Octamer/Sox element was sufficient for inducing the proper expression of a green fluorescent protein reporter gene even in human embryonic stem (ES) cells. The potential of OCT4 and SOX2 to bind to this element was verified by electrophoretic mobility shift assays with extracts from F9 embryonal carcinoma cells and embryonic germ cells derived from embryonic day 12.5 embryos. However, in ES cell extracts, a complex of OCT4 with an undefined factor preferentially bound to the Octamer/Sox element. Thus, Nanog transcription may be regulated through an interaction between Oct4 and Sox2 or a novel pluripotential cell-specific Sox element-binding factor which is prominent in ES cells.
|
Volume |
25(6)
|
Pages |
2475-85
|
Published |
2005-3-1
|
DOI |
10.1128/MCB.25.6.2475-2485.2005
|
PII |
25/6/2475
|
PMID |
15743839
|
PMC |
PMC1061601
|
MeSH |
5' Flanking Region / genetics
Animals
Base Sequence
Binding Sites / genetics
DNA-Binding Proteins / genetics*
DNA-Binding Proteins / metabolism
DNA-Binding Proteins / physiology*
Electrophoretic Mobility Shift Assay
Embryo, Mammalian / cytology
Gene Expression Regulation*
Genes, Reporter / genetics
Green Fluorescent Proteins / analysis
Green Fluorescent Proteins / genetics
HMGB Proteins
Homeodomain Proteins / genetics*
Humans
Luciferases / analysis
Luciferases / genetics
Mice
Molecular Sequence Data
Mutation / genetics
Nanog Homeobox Protein
Nuclear Proteins / genetics
Nuclear Proteins / metabolism
Nuclear Proteins / physiology*
Octamer Transcription Factor-3
Response Elements / genetics*
SOXB1 Transcription Factors
Sequence Deletion / genetics
Stem Cells / metabolism*
Transcription Factors / genetics
Transcription Factors / metabolism
Transcription Factors / physiology*
Transcription, Genetic
|
IF |
3.611
|
Times Cited |
361
|
WOS Category
|
BIOCHEMISTRY & MOLECULAR BIOLOGY
CELL BIOLOGY
|
Resource |
Human embryonic stem cells |
KhES-1 |