The regulation of the UCH-L1 gene by transcription factor NF-κB in podocytes

Cell Signal. 2013 Jul;25(7):1574-85. doi: 10.1016/j.cellsig.2013.03.018. Epub 2013 Apr 6.

Abstract

In kidney, the ubiquitin carboxy-terminal hydrolase 1 (UCH-L1) is involved in podocyte injury and proteinuria but details of the mechanism underlying its regulation are not known. Activation of NF-κB is thought to be the predominant risk factor for kidney disease; therefore, it is postulated that UCH-L1 may be one of the NF-κB target genes. In this study, we investigated the involvement of NF-κB activation in the regulation of UCH-L1 expression and the function of murine podocytes. Stimulation of podocytes with the cytokines TNF-α and IL-1β up-regulated UCH-L1 expression rapidly at the mRNA and protein levels and the NF-κB-specific inhibitor pyrrolidine dithiocarbamate resulted in down-regulation. NF-κB up-regulates UCH-L1 via binding the --300 bp and --109 bp sites of its promoter, which was confirmed by the electrophoretic mobility shift assay of DNA-nuclear protein binding. In the renal biopsy from lupus nephritis patients, the expressions of NF-κB and UCH-L1 increased in immunohistochestry staining and were positively correlated. Activation of NF-κB up-regulates UCH-L1 expression following changing of other podocytes molecules, such as nephrin and snail. These results suggest that activation of the NF-κB signaling pathway could be the major pathogenesis to up-regulate UCH-L1 in podocyte injury, followed by the turnover of other molecules, which might result in morphological changes and dysfunction of podocytes. This work help us to understand the effect of NF-κB on specific target molecules of podocytes, and suggest that targeting the NF-κB-UCH-L1 interaction could be a novel therapeutic strategy for the treatment of podocyte lesions and proteinuria.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Animals
  • Base Sequence
  • Binding Sites
  • Consensus Sequence
  • Genes, Reporter
  • HEK293 Cells
  • Humans
  • Interleukin-1beta / physiology
  • Kidney Glomerulus / metabolism
  • Luciferases, Renilla / biosynthesis
  • Luciferases, Renilla / genetics
  • Lupus Nephritis / metabolism
  • Lupus Nephritis / pathology
  • Membrane Proteins / metabolism
  • Mice
  • Podocytes / metabolism*
  • Promoter Regions, Genetic
  • RNA, Messenger / genetics
  • RNA, Messenger / metabolism
  • Snail Family Transcription Factors
  • Transcription Factor RelA / metabolism*
  • Transcription Factors / metabolism
  • Transcriptional Activation*
  • Tumor Necrosis Factor-alpha / physiology
  • Ubiquitin Thiolesterase / genetics*
  • Ubiquitin Thiolesterase / metabolism

Substances

  • Interleukin-1beta
  • Membrane Proteins
  • RNA, Messenger
  • Rela protein, mouse
  • Snail Family Transcription Factors
  • Transcription Factor RelA
  • Transcription Factors
  • Tumor Necrosis Factor-alpha
  • Ubiquitin carboxyl-Terminal Hydrolase L-1, mouse
  • nephrin
  • Luciferases, Renilla
  • Ubiquitin Thiolesterase