Isolation, characterization and chromosomal localization of the human GADD153 gene

Abstract

We report the isolation and characterization of the growth arrest and DNA-damage-inducible gene, GADD153, from human cells and show that it is localized in the region 12q13.1-q13.2 on chromosome 12. Comparison of the human gene with the previously described hamster gene revealed a high level of conservation in both the structural and regulatory regions of the genes. Each is composed of four exons with intron/exon junctions maintained at the identical positions. The human Gadd153 protein shares 91% identity with the hamster protein in amino acid sequence, and 78% identity in nucleotide sequence. A 900-bp fragment of 5' flanking sequence from the human gene, when linked to the bacterial cat reporter gene, was found to exhibit promoter activity in HeLa cells which could be further activated by treatment with the DNA alkylating agent, methyl methanesulfonate. Sequence analysis indicated that the human promoter region is relatively G+C-rich and contains putative binding sites for multiple transcription factors, including recognition sites for TATA- and CAAT-binding proteins, six Sp1-binding sites, an activator protein-1 binding site, an E-26-specific sequence-binding protein-1 DNA-binding site, and four interleukin-6 response elements. Many of these sites are also present in an identical position in the hamster gene suggesting they may play an important role in regulating GADD153 expression.

Characteristic sequence motifs at the breakpoints of the hybrid genes FUS/CHOP, EWS/CHOP and FUS/ERG in myxoid liposarcoma and acute myeloid leukemia

“…Sequence analysis of the entire PCR fragment revealed that the ®rst intron of CHOP was 2669 bp with 50% A+T composition (A: 717; T: 613) and contained two Alu repeat sequences, one of the Sx-type and one of the Sc-type (Figure 1). There was, however, a discrepancy between the present and the previously published data (Park et al, 1992) as regards the size of intron 1. In our material, CHOP intron 1 was 2669 bp, whereas it was previously estimated to be approximately 1500 bp.…”

“…To verify whether or not the PCR product represented intron 1 of the CHOP gene, the fragment was puri®ed and analysed by direct cycling sequencing using the inner primers CHOP53F and CHOP105R and the sequence was compared with the known CHOP sequence (Park et al, 1992). Using CHOP53F as primer, the last 17 bases of exon 1 as well as the ®rst 10 bases of introl 1 (with only one base substitution compared to the known CHOP sequence) were identi®ed.…”

“…All primers were based on the full length cDNA sequences of CHOP (Park et al, 1992), ERG (Reddy et al, 1987), EWS (Delattre et al, 1992) and FUS (Crozat et al, 1993).…”

CCAAT/Enhancer-binding protein homologous protein (CHOP) decreases bone formation and causes osteopenia

“…CCAAT/enhancer binding proteins (C/EBP) are a family of nuclear proteins, which include C/EBP homologous protein (CHOP), also termed growth arrest and DNA damage-inducible gene (GADD) 153, or DNA damage inducible transcript 3 (DDIT3) [1][2][3][4]. C/EBP proteins contain highly conserved DNA-binding and leucine dimerization domains, and can form homoand hetero-dimers that bind to similar sequence motifs.…”

A supramicromolar elevation of intracellular free calcium ([Ca2+]i) is consistently required to induce the execution phase of apoptosis

“…This promoter sequence has previously been used to monitor gadd153 up-regulation, in particular after stimulation by agents that increase [Ca 2+ ] i such as ionomycin. 42,43 A series of experiments with agents that directly elevate [Ca 2+ ] i demonstrated that, in TSU and NRP152 stably transfected with pGadd153 ± EGFP-1, gadd153 RNA expression could be monitored in cells by measuring the amount of EGFP fluorescence. In individual cells, green fluorescence and morphological changes were monitored using the longitudinal spectrofluorometric method previously validated for [Ca 2+ ] i measurements and adjusted for GFP wavelength.…”