Proximal Promoter Sequences Mediate Cell-specific and Elevated Expression of the Favorable Prognosis Marker TrkA in Human Neuroblastoma Cells

Chang, Baochong B.; Persengiev, Stephan P.; Diego, Juana Gutiérrez de; Sacristán, María P.; Zanca, Dionisio Martin; Kilpatrick, Daniel L.

doi:10.1074/jbc.273.1.39

Cited by 14 publications

(20 citation statements)

References 44 publications

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“…Disruption of this sequence or mutations that destroyed the dyad symmetry drastically reduced transcription activity to levels similar to those of inactive fusions. A similar motif is conserved at an equivalent position in the human trkA promoter, although in this case there are four G residues separating the three-nucleotide inverted repeat (Chang et al, 1998). This mouse trkA cisregulatory element was capable of binding nuclear Gel mobility shift assays performed using N2a nuclear extracts and the 771/753 double-stranded oligonucleotide as probe, in the presence or absence of 2 mg of anti-Sp1, anti-AP2 or anti-Egr1 antibodies (Santa Cruz Biotechnology, Santa Cruz, CA, USA).…”

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confidence: 98%

“…unrelated, oligonucleotide; M: mutated oligonucleotide consensus sequences for transcription factors; in particular the segment between 793 and 756 contains three potential binding sites for Sp1, two for AP-2, and several for GCF (Figure 5a). Most of these sites are conserved in the promoter region of human trkA (Chang et al, 1998). The 771/753 oligonucleotide contains consensus binding sites for Sp1, AP-2 and, with a single mismatch, for Egr-1.…”

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confidence: 99%

“…In contrast, the human trkA gene seems to initiate transcription at several sites, all close to the AUG codon. The reason for this di erence is not clear; however, it is possible that di erent transcription start sites are used in di erent cell types; thus, we have de®ned the mouse trkA start site in RNA from primary embryonic DRGs, while that of the human gene was determined in transformed cell lines (Chang et al, 1998).…”

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confidence: 99%

“…Antibodies were added to the reaction mixture and incubated for 1 ± 2 h on ice before adding the labeled probe. I, II and III indicate DNA-protein complexes; SS indicates a supershifted complex formed in the presence of anti-Sp1 antibody proteins present in trkA-expressing cell lines, giving rise to three speci®c DNA/protein complexes, which resembled those observed in human neuroblastoma cell lines, using a longer human DNA probe (Chang et al, 1998). These complexes were undetectable in trkA non-expressing cell lines, suggesting that the nuclear factors involved in their formation may contribute to the tissue-speci®c expression of trkA.…”

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Molecular cloning and characterization of the 5′ region of the mouse trkA proto-oncogene

et al. 1999

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The trkA proto-oncogene encodes a high-a nity NGF receptor that is essential for the survival, di erentiation and maintenance of many neural and non-neural cell types. Altered expression of the trkA gene or trkA receptor malfunction have been implicated in neurodegeneration, tumor progression and oncogenesis. We have cloned and characterized the 5' region of the mouse trkA gene and have identi®ed its promoter. trkA promoter sequences are GC-rich, lack genuine TATA or CAAT boxes, and are contained within a CpG island which extends over the entire ®rst coding exon. The mouse trkA transcription start site is located 70/71 bp upstream to the AUG translation initiation codon. Sequence analysis showed that the gene encoding the insulin receptor-related receptor, IRR, is located just 1.6 kbp upstream to the trkA gene and is transcribed in the opposite direction. We have used trkA-CAT transcriptional fusions to study trkA promoter function in transient transfection experiments. RNase protection assays and CAT protein ELISA analyses showed that a 150 bp long DNA segment, immediately upstream to the start site, is su cient to direct accurate transcription in trkA-expressing cells. Dissection of this fragment allowed us to identify a 13 bp cis-regulatory element essential for both promoter activity and cell-type speci®c expression. Deletion of this 13 bp segment as well as modi®cation of its sequence by site-directed mutagenesis led to a dramatic decline in promoter activity. Gel mobility shift assays carried out with double-stranded oligonucleotides containing the 13 bp element revealed several speci®c DNA-protein complexes when nuclear extracts from trkA-expressing cells were used. Supershift experiments showed that the Sp1 transcription factor was a component of one of these complexes. Our results identify a minimal trkA gene promoter, located very close to the transcription start site, and de®ne a 13 bp enhancer within this promoter sequence.

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Molecular cloning and characterization of the 5′ region of the mouse trkA proto-oncogene

et al. 1999

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“…The genomic region of 1085 bp upstream from the 5' end of the hb-1 cDNA clone was analysed ( Figure 2). In addition to the high CG content, the sequence analysis revealed the absence of TATA box and the presence of multiple putative recognition sites for transcription factors, including Sp1, AP2, GC and ATF, which have also been reported in the proximal regulatory region of the trkA gene (Chang et al, 1998;Greco et al, 1996). Besides AP2, other transcription factors known to be expressed in brain, such as BRN2, or both in heart and brain, such as AML1 and Nkx2.5, also have recognition sites in this region (Figure 2) (Heinemeyer et al, 1998;Quandt et al, 1995).…”

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confidence: 99%

Genomic characterization of the human trkC gene

et al. 1998

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The trkC gene encodes the high-anity receptor for neurotrophin 3 and plays an important role in the regulation of the survival and dierentiation of the mammalian nervous system and in heart development. Chromosomal rearrangements of trkC have been recently reported in congenital ®brosarcoma and it has been proposed that abnormal activation of this gene might be involved in tumor development. To facilitate the search for new mutations and rearrangements in the human trkC locus we have partially characterized its genomic organization by restriction mapping and have obtained the complete intron-exon structure. Our results show that human trkC consists of 20 exons, including two that encode the inserts present in the extracellular and tyrosine kinase domains, and another two that encode the carboxyl-terminal tail of the truncated TRKC isoform. Analysis of the 5'¯anking region revealed the absence of TATA box, a very high content in C/G compatible with a CpG island and the presence of putative binding sites for the AP1, AP2, GC, ATF, BRN2, AML1 and Nkx2.5 transcription factors.

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Gene Expression of Neurotrophins and Their Receptors in Cultured Rat Vascular Smooth Muscle Cells

Nemoto

Fukamachi

Nemoto

et al. 1998

Biochemical and Biophysical Research Communications

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Proximal Promoter Sequences Mediate Cell-specific and Elevated Expression of the Favorable Prognosis Marker TrkA in Human Neuroblastoma Cells

Cited by 14 publications

References 44 publications

Molecular cloning and characterization of the 5′ region of the mouse trkA proto-oncogene

Molecular cloning and characterization of the 5′ region of the mouse trkA proto-oncogene

Genomic characterization of the human trkC gene

Gene Expression of Neurotrophins and Their Receptors in Cultured Rat Vascular Smooth Muscle Cells

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