GADD45a promoter regulation by a functional genetic variant associated with acute lung injury

Sumegha Mitra, Michael S. Wade, Xiaoguang Sun, Nurgul Moldobaeva, Carlos Flores, Ma Shwu-Fan, Wei Zhang, Joe G.N. Garcia, Jeffrey R. Jacobson

Research output: Contribution to journalArticle

8 Scopus citations

Abstract

Rationale: Growth arrest DNA damage inducible alpha (GADD45a) is a stress-induced gene we have shown to participate in the pathophysiology of ventilator-induced lung injury (VILI) via regulation of mechanical stress-induced Akt ubiquitination and phosphorylation. The regulation of GADD45a expression by mechanical stress and its relationship with acute lung injury (ALI) susceptibility and severity, however, remains unknown. Objectives: We examined mechanical stress-dependent regulatory elements (MSRE) in the GADD45a promoter and the contribution of promoter polymorphisms in GADD45a expression and ALI susceptibility. Methods and Results: Initial studies in GADD45a knockout and heterozygous mice confirmed the relationship of GADD45a gene dose to VILI severity. Human lung endothelial cells (EC) transfected with a luciferase vector containing the full length GADD45a promoter sequence (-771 to +223) demonstrated a >4 fold increase in GADD45a expression in response to 18% cyclic stretch (CS, 4 h) compared to static controls while specific promoter regions harboring CS-dependent MSRE were identified using vectors containing serial deletion constructs of the GADD45a promoter. In silico analyses of GADD45a promoter region (-371 to -133) revealed a potential binding site for specificity protein 1 (SP1), a finding supported by confirmed SP1 binding with the GADD45a promoter and by the significant attenuation of CS-dependent GADD45a promoter activity in response to SP1 silencing. Separately, case-control association studies revealed a significant association of a GADD45a promoter SNP at -589 (rs581000, G>C) with reduced ALI susceptibility. Subsequently, we found allelic variation of this SNP is associated with both differential GADD45a expression in mechanically stressed EC (18% CS, 4 h) and differential binding site of interferon regulatory factor 7 (IRF7) at this site. Conclusion: These results strongly support a functional role for GADD45a in ALI/VILI and identify a specific gene variant that confers risk for ALI.

Original languageEnglish (US)
Article numbere100169
JournalPloS one
Volume9
Issue number6
DOIs
StatePublished - Jun 18 2014
Externally publishedYes

ASJC Scopus subject areas

  • Biochemistry, Genetics and Molecular Biology(all)
  • Agricultural and Biological Sciences(all)
  • General

Fingerprint Dive into the research topics of 'GADD45a promoter regulation by a functional genetic variant associated with acute lung injury'. Together they form a unique fingerprint.

  • Cite this

    Mitra, S., Wade, M. S., Sun, X., Moldobaeva, N., Flores, C., Shwu-Fan, M., Zhang, W., Garcia, J. G. N., & Jacobson, J. R. (2014). GADD45a promoter regulation by a functional genetic variant associated with acute lung injury. PloS one, 9(6), [e100169]. https://doi.org/10.1371/journal.pone.0100169