Dataset Information

Characterization of the mouse transforming growth factor-beta 1 promoter and activation by the Ha-ras oncogene.

ABSTRACT: We have cloned and sequenced a mouse genomic transforming growth factor beta 1 (TGF-beta 1) DNA fragment that includes the 5' untranslated and regulatory regions of the gene. High-sequence homology with the human TGF-beta 1 gene (66% nucleotide identity in 2.7 kb of DNA upstream of the translational start site) suggested evolutionary conservation of transcriptional regulation for TGF-beta 1. The absence of TATA or CAAT box sequences but the presence of several Sp1-binding and AP-2-like sequences in the promoter region was noted, as previously reported for the human gene. Two transcriptional initiation sites separated by 290 bp were identified by S1 nuclease analysis; these corresponded to transcripts with 866 and 576 nucleotides of 5' untranslated leader sequence. S1 analysis of different mouse tissues indicated that the two transcripts were present in the same ratio even though the total level of TGF-beta 1 mRNA transcripts varied between tissues. Promoter activity adjacent to both transcriptional start sites was demonstrated by using chloramphenicol acetyltransferase fusion genes assayed in mouse AKR-2B fibroblast cells. Transcriptional activation of the promoter by the Ha-ras oncogene was also demonstrated. The minimal promoter constructs (113 and 104 bp 5' of the first and second transcriptional start sites, respectively) were sufficient for induction by Ha-ras. These studies characterize the 5' structure and basal promoter activity of the mouse TGF-beta 1 gene as well as the transcriptional activation of TGF-beta 1 by the Ha-ras oncogene.

SUBMITTER: Geiser AG

PROVIDER: S-EPMC359593 | biostudies-other | 1991 Jan

REPOSITORIES: biostudies-other

ACCESS DATA

Publications

Characterization of the mouse transforming growth factor-beta 1 promoter and activation by the Ha-ras oncogene.

Geiser A G AG Kim S J SJ Roberts A B AB Sporn M B MB

Molecular and cellular biology 19910101 1

We have cloned and sequenced a mouse genomic transforming growth factor beta 1 (TGF-beta 1) DNA fragment that includes the 5' untranslated and regulatory regions of the gene. High-sequence homology with the human TGF-beta 1 gene (66% nucleotide identity in 2.7 kb of DNA upstream of the translational start site) suggested evolutionary conservation of transcriptional regulation for TGF-beta 1. The absence of TATA or CAAT box sequences but the presence of several Sp1-binding and AP-2-like sequences ...[more]

PMID: 1986255

Similar Datasets

Project description:Variations in the Toll-interacting protein (TOLLIP) gene have been identified in genome-wide association studies to correlate with risk of disease, mortality, and response to N-acetylcysteine therapy in idiopathic pulmonary fibrosis. Although TOLLIP is known to modulate innate immune responses, its relevance in organ fibrogenesis remains unknown. Prior work in the literature suggests TOLLIP dampens transforming growth factor beta (TGFβ) signaling in human cell lines. In this study, we examined the role of TOLLIP in mouse lung fibroblast (MLF) responses to TGFβ and in the bleomycin model of experimental lung fibrosis using Tollip-/- mice. We hypothesize that if TOLLIP negatively regulates TGFβ signaling, then Tollip-/- mouse lung fibroblasts (MLFs) would have enhanced response to TGFβ treatment, and Tollip-/- mice would develop increased fibrosis following bleomycin challenge. Primary MLFs were stimulated with TGFβ (1 ng/mL) for 24 h. RNA was obtained to assess global transcriptional responses by RNA-seq and markers of myofibroblast transition by qPCR. Functional assessment of TGFβ-stimulated MLFs included cell migration by scratch assay, cell proliferation, and matrix invasion through Matrigel. In the in vivo model of lung fibrosis, Tollip-/- mice and wild-type (WT) littermates were administered bleomycin intratracheally and assessed for fibrosis. We further examined TGFβ signaling in vivo after bleomycin injury by SMAD2, ERK1/2, and TGFβR1 Western blot. In response to TGFβ treatment, both WT and Tollip-/- MLFs exhibited global transcriptional changes consistent with myofibroblast differentiation. However, Tollip-/- MLFs showed greater number of differentially expressed genes compared to WT MLFs and greater upregulation of Acta2 by qPCR. Functionally, Tollip-/- MLFs also exhibited increased migration and Matrigel invasiveness compared to WT. We found evidence of enhanced TGFβ signaling in Tollip-/- through SMAD2 in vitro and in vivo. Tollip-/- mice experienced lower survival using a standard weight-adjusted dosing without evidence of differences in fibrosis at Day 21. With adjustment of dosing for sex, no differences were observed in fibrosis at Day 21. However, Tollip-/- mice had greater weight loss and increased bronchoalveolar lavage fluid total protein during early resolution at Day 14 compared to WT without evidence of differences in acute lung injury at Day 7, suggesting impaired resolution of lung injury.

Dataset Information

Characterization of the mouse transforming growth factor-beta 1 promoter and activation by the Ha-ras oncogene.

Publications

Characterization of the mouse transforming growth factor-beta 1 promoter and activation by the Ha-ras oncogene.

Similar Datasets

OmicsDI is part of the ELIXIR infrastructure

Tweets