scholarly journals Transcriptional modulation of the pre-implantation embryo-specific Rnf35 gene by the Y-box protein NF-Y/CBF

2005 ◽  
Vol 387 (2) ◽  
pp. 367-375 ◽  
Author(s):  
Chiu-Jung HUANG ◽  
Shinn-Chih WU ◽  
Kong-Bung CHOO
Keyword(s):  
Chinese Hamster Ovary ◽  
Core Promoter ◽  
Chinese Hamster ◽  
Ring Finger ◽  
Early Embryo ◽  
Promoter Element ◽  
Mobility Shift ◽  
Core Promoter Element ◽  
Stage Of Development ◽  
Transcriptional Modulation

Maternal-to-zygotic transition of a fertilized egg and the subsequent pre-implantation development of the embryo involve zygotic genome activation and reprogramming of gene expression. The goal of the present study is to establish a model suitable for the characterization of transcriptional modulation of mammalian pre-implantation development. Rnf35 is a mouse RING-finger protein gene that is temporally transcribed in the early embryo, but is permanently silenced before the blastocyst stage of development. We first show that the Chinese-hamster ovary-K1 cells are unique in supporting Rnf35 promoter activities in transient transfection assays. Using the permissive Chinese-hamster ovary-K1 cell line, we show that Rnf35 transcription is driven by an Inr (initiator) core promoter element in the absence of a TATA box; the Inr promoter function is confirmed by direct microinjection of mouse one-cell embryos. This is the first demonstration of the involvement of an Inr core promoter element in transcription in pre-implantation development. We show that the Rnf35 promoter is regulated by three obligatory Y-box (CCAAT-box) elements: two Y boxes (YI and YII) located at −81 are coupled in a palindrome and act synergistically in contributing to Rnf35 transcription; the third Y box (YIII) is situated at −13, just upstream of the Inr element, and may be an integral part of the Inr function. Electrophoretic mobility-shift assays and competition experiments further reveal that the YI box is bound by the ubiquitous NF-Y (nuclear factor-Y)/CBF (CCAAT-binding factor) and that YII is targeted by an unidentified protein(s) that acts synergistically with the NF-Y. We suggest that the NF-Y, targeting at a Y-box sequence, may function as an important activator in transcriptional regulation of the Rnf35 gene in the pre-implantation embryo.

scholarly journals Two Inr Elements Are Important for Mediating the Activity of the Proximal Promoter of the Human Gonadotropin-Releasing Hormone Receptor Gene

Endocrinology ◽  
2003 ◽  
Vol 144 (2) ◽  
pp. 518-527 ◽  
Author(s):  
Ruby L. C. Hoo ◽  
Elly S. W. Ngan ◽  
Peter C. K. Leung ◽  
Billy K. C. Chow
Keyword(s):  
Core Promoter ◽  
Receptor Gene ◽  
Fold Increase ◽  
Gnrh Receptor ◽  
Proximal Promoter ◽  
Dependent Manner ◽  
Promoter Element ◽  
Mobility Shift ◽  
Core Promoter Element ◽  
The Core

Differential usage of several transcription start sites in the human GnRH receptor gene was evident in human brain and pituitary. To locate the promoter responsible for a cluster of the 3′ CAP sites from −635 to −578 (relative to ATG) found in the pituitary, a proximal promoter element was identified at −677/−558 by 5′ and 3′ deletion mutant analysis. The promoter element drove a 13.1 ± 0.6-fold increase in reporter gene activity in an orientation-dependent manner in the mouse gonadotrope-derived αT3–1 cells. Within the core promoter element, two functional AT-rich Inr motifs, interacting with the same protein factor with different affinities, were identified. By Southwestern blot analysis and competitive gel mobility shift assays, multiple nuclear factors (36–150 kDa) were found to interact specifically with the core promoter element. Interestingly, these nuclear proteins also interacted with a previously identified distal promoter of the human GnRH receptor gene. Taken together, our studies suggested that these two promoters share common protein factors to regulate transcription initiations at two different regions. Additional mechanisms are needed to modulate the efficiencies of individual promoters for developmental and/or tissue-specific regulations.

scholarly journals Core Promoter Structure in the Oomycete Phytophthora infestans

2004 ◽  
Vol 3 (1) ◽  
pp. 91-99 ◽  
Author(s):  
Adele McLeod ◽  
Christine D. Smart ◽  
William E. Fry
Keyword(s):  
Phytophthora Infestans ◽  
Promoter Region ◽  
Core Promoter ◽  
Cell Protein ◽  
Promoter Element ◽  
Core Promoter Region ◽  
Mobility Shift ◽  
Promoter Structure ◽  
Core Promoter Element ◽  
The Core

ABSTRACT We have investigated the core promoter structure of the oomycete Phytophthora infestans. The transcriptional start sites (TSS) of three previously characterized P. infestans genes, Piexo1, Piexo3, and Piendo1, were determined by primer extension analyses. The TSS regions were homologous to a previously identified 16-nucleotide (nt) core sequence that overlaps the TSS in most oomycete genes. The core promoter regions of Piexo1 and Piendo1 were investigated by using a transient protoplast expression assay and the reporter gene β-glucuronidase. Mutational analyses of the promoters of Piexo1 and Piendo1 showed that there is a putative core promoter element encompassing the TSS (−2 to + 5) that has high sequence and functional homology to a known core promoter element present in other eukaryotes, the initiator element (Inr). Downstream and flanking the Inr is a highly conserved oomycete promoter region (+7 to + 15), hereafter referred to as FPR (flanking promoter region), which is also important for promoter function. The importance of the 19-nt core promoter region (Inr and FPR) in Piexo1 and Piendo1 was further investigated through electrophoretic mobility shift assays (EMSA). The EMSA studies showed that (i) both core promoters were able to specifically bind a protein or protein complex in a P. infestans whole-cell protein extract and (ii) the same mutations that reduced binding of the EMSA complex also reduced β-glucuronidase (GUS) levels in transient expression assays. The consistency of results obtained using two different assays (GUS transient assays [in vivo] and EMSA studies [in vitro]) supports a convergence of inference about the relative importance of specific nucleotides within the 19-nt core promoter region.

Association of Variants in Critical Core Promoter Element of Angiotensinogen Gene With Increased Risk of Essential Hypertension in Japanese

Hypertension ◽  
1997 ◽  
Vol 30 (3) ◽  
pp. 321-325 ◽  
Author(s):  
Noriyuki Sato ◽  
Tomohiro Katsuya ◽  
Hiromi Rakugi ◽  
Seiju Takami ◽  
Yukiko Nakata ◽  
...  
Keyword(s):  
Essential Hypertension ◽  
Core Promoter ◽  
Promoter Element ◽  
Core Promoter Element ◽  
Angiotensinogen Gene ◽  
Increased Risk

scholarly journals DTIE, a novel core promoter element that directs start site selection in TATA-less genes

2015 ◽  
Vol 44 (3) ◽  
pp. 1080-1094 ◽  
Author(s):  
Nadav Marbach-Bar ◽  
Anat Bahat ◽  
Shaked Ashkenazi ◽  
Michal Golan-Mashiach ◽  
Ora Haimov ◽  
...  
Keyword(s):  
Site Selection ◽  
Core Promoter ◽  
Start Site ◽  
Promoter Element ◽  
Core Promoter Element

The DPE, a Conserved Downstream Core Promoter Element That Is Functionally Analogous to the TATA Box

1998 ◽  
Vol 63 (0) ◽  
pp. 75-82 ◽  
Author(s):  
T.W. BURKE ◽  
P.J. WILLY ◽  
A.K. KUTACH ◽  
J.E.F. BUTLER ◽  
J.T. KADONAGA
Keyword(s):  
Core Promoter ◽  
Tata Box ◽  
Promoter Element ◽  
Core Promoter Element

scholarly journals Identification of a novel p53 promoter element involved in genotoxic stress-inducible p53 gene expression.

1995 ◽  
Vol 15 (8) ◽  
pp. 4489-4496 ◽  
Author(s):  
X Sun ◽  
H Shimizu ◽  
K Yamamoto
Keyword(s):  
Stress Response ◽  
Anticancer Drugs ◽  
Promoter Activity ◽  
Core Promoter ◽  
Genotoxic Stress ◽  
Promoter Element ◽  
Promoter Activation ◽  
Nf Kappa B ◽  
Core Promoter Element ◽  
Genotoxic Agents

p53 is recruited in response to DNA-damaging genotoxic stress and plays an important role in maintaining the integrity of the genome. We show that exposure of cells to various genotoxic agents, including anticancer drugs such as mitomycin and 5-fluorouracil, results in an increase in p53 mRNA levels and in p53 promoter activation, indicating that the p53 genotoxic stress response is partly regulated at the transcriptional level. The results of the p53 promoter analysis show that a novel p53 promoter element, termed a p53 core promoter element (from -70 to -46), is essential for basal p53 promoter activity and promoter activation induced by genotoxic agents such as anticancer drugs and UV. Although a kappa B motif partially overlaps with this element and those genotoxic agents activate NF-kappa B, it does not play a major role in p53 genotoxic stress response: NF-kappa B p65 expression did not induce significant p53 promoter activation, and NF-kappa B inhibitors (N-acetyl cysteine and I kappa B alpha) did not inhibit genotoxic stress-inducible p53 promoter activation. Finally, we characterized nuclear factors, the binding of which to the p53 core promoter element is essential for basal p53 promoter activity and p53 promoter activation induced by genotoxic agents.

The Initiator Motif is Preferentially Used as the Core Promoter Element in Ionizing Radiation-Responsive Genes

2006 ◽  
Vol 166 (5) ◽  
pp. 810-813 ◽  
Author(s):  
Jianyu Wu ◽  
Kazuhiro Daino ◽  
Sachiko Ichimura ◽  
Mitsuru Nenoi
Keyword(s):  
Ionizing Radiation ◽  
Core Promoter ◽  
Promoter Element ◽  
Core Promoter Element ◽  
The Core

scholarly journals Isolation and characterization of the rat huntingtin promoter

1998 ◽  
Vol 336 (1) ◽  
pp. 227-234 ◽  
Author(s):  
Carsten HOLZMANN ◽  
Winfried MÄUELER ◽  
Dirk PETERSOHN ◽  
Thorsten SCHMIDT ◽  
Gerald THIEL ◽  
...  
Keyword(s):  
Protein Interactions ◽  
Binding Sites ◽  
Neurodegenerative Disorder ◽  
Core Promoter ◽  
Chinese Hamster ◽  
Gc Content ◽  
Ovary Cell ◽  
Mobility Shift ◽  
Isolation And Characterization ◽  
Hd Gene

Huntington's disease (HD) is a neurodegenerative disorder caused by a (CAG)>37 repeat expansion in a novel gene of unknown function. Although the huntingtin gene is expressed in neuronal and non-neuronal tissues, the disease affects nerve cells of selected regional areas of the central nervous system. To gain insight into the regulation of the HD gene we analysed 1348 bp of the rat huntingtin promoter region. This region lacks a TATA and a CAAT box, is rich in GC content and has several consensus sequences for binding sites for SP1, PEA3, Sif and H2A. The stretch between nucleotides -56 and -206 relative to the first ATG is highly conserved between human and rodents and it harbours several potential binding sites for transcription factors. We analysed deletion mutants fused with the chloramphenicol acetyltransferase reporter gene in transfected, HD-expressing neuronal (NS20Y, NG108-15) and non-neuronal Chinese hamster ovary cell lines. Hence these cells should contain the required trans-acting factors necessary for HD gene expression. Partial deletion of the evolutionarily conserved part of the promoter significantly decreases the activity in both neuronal and non-neuronal cells, indicating that the core promoter activity is located between nucleotides -332 and -15. DNase I footprinting and electrophoretic mobility-shift assays were used to define the nucleotide positions and binding affinity of DNA–protein interactions.

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