Novel RNA-binding activity of NQO1 promotes SERPINA1 mRNA translation

Free Radic Biol Med. 2016 Oct;99:225-233. doi: 10.1016/j.freeradbiomed.2016.08.005. Epub 2016 Aug 8.

Abstract

NAD(P)H: quinone oxidoreductase (NQO1) is essential for cell defense against reactive oxidative species, cancer, and metabolic stress. Recently, NQO1 was found in ribonucleoprotein (RNP) complexes, but NQO1-interacting mRNAs and the functional impact of such interactions are not known. Here, we used ribonucleoprotein immunoprecipitation (RIP) and microarray analysis to identify comprehensively the subset of NQO1 target mRNAs in human hepatoma HepG2 cells. One of its main targets, SERPINA1 mRNA, encodes the serine protease inhibitor α-1-antitrypsin, A1AT, which is associated with disorders including obesity-related metabolic inflammation, chronic obstructive pulmonary disease (COPD), liver cirrhosis and hepatocellular carcinoma. Biotin pulldown analysis indicated that NQO1 can bind the 3' untranslated region (UTR) and the coding region (CR) of SERPINA1 mRNA. NQO1 did not affect SERPINA1 mRNA levels; instead, it enhanced the translation of SERPINA1 mRNA, as NQO1 silencing decreased the size of polysomes forming on SERPINA1 mRNA and lowered the abundance of A1AT. Luciferase reporter analysis further indicated that NQO1 regulates SERPINA1 mRNA translation through the SERPINA1 3'UTR. Accordingly, NQO1-KO mice had reduced hepatic and serum levels of A1AT and increased activity of neutrophil elastase (NE), one of the main targets of A1AT. We propose that this novel mechanism of action of NQO1 as an RNA-binding protein may help to explain its pleiotropic biological effects.

Keywords: A1AT; NQO1; Neutrophil elastase; RNA binding protein; SERPINA1; mRNA translation.

Publication types

  • Research Support, N.I.H., Intramural

MeSH terms

  • Animals
  • Binding Sites
  • Gene Expression Regulation
  • Genes, Reporter
  • Hep G2 Cells
  • Humans
  • Leukocyte Elastase / genetics
  • Leukocyte Elastase / metabolism
  • Luciferases / genetics
  • Luciferases / metabolism
  • Mice
  • Mice, Inbred C57BL
  • Mice, Knockout
  • Microarray Analysis
  • NAD(P)H Dehydrogenase (Quinone) / antagonists & inhibitors
  • NAD(P)H Dehydrogenase (Quinone) / genetics*
  • NAD(P)H Dehydrogenase (Quinone) / metabolism
  • Protein Binding
  • Protein Biosynthesis*
  • RNA, Messenger / genetics*
  • RNA, Messenger / metabolism
  • RNA, Small Interfering / genetics
  • RNA, Small Interfering / metabolism
  • Ribonucleoproteins / genetics
  • Ribonucleoproteins / metabolism
  • Signal Transduction
  • alpha 1-Antitrypsin / genetics*
  • alpha 1-Antitrypsin / metabolism

Substances

  • RNA, Messenger
  • RNA, Small Interfering
  • Ribonucleoproteins
  • SERPINA1 protein, human
  • alpha 1-Antitrypsin
  • Luciferases
  • NAD(P)H Dehydrogenase (Quinone)
  • NQO1 protein, human
  • Leukocyte Elastase