Identification of an H2 O2 permeable PIP aquaporin in barley and a serine residue promoting H2 O2 transport

Physiol Plant. 2017 Jan;159(1):120-128. doi: 10.1111/ppl.12508. Epub 2016 Oct 2.

Abstract

A barley (Hordeum vulgare) plasma membrane type aquaporin, HvPIP2;5, was identified as an H2 O2 permeable aquaporin among 21 barley and rice PIPs examined in the heterologous expression system using Saccharomyces cerevisiae. Four TIPs were also detected as H2 O2 -transporting aquaporins among 15 barley and rice TIPs. Influx of H2 O2 into yeast cells expressing HvPIP2;5 was determined with a florescent-dye-based assay. Indirect immunofluorescence indicated that the expression of HvPIP2;5 protein was ubiquitous in root tissues, and was also weakly observed in leaf epidermal cells and cells in the vascular bundle. Point mutated variants of HvPIP2;5 were generated by the site-directed mutagenesis. Growth assays of yeast cells expressing these mutated HvPIP2;5 proteins suggested that Ser-126 in HvPIP2;5 has a large impact on H2 O2 transport with a minor influence on the HvPIP2;5-mediated water transport.

MeSH terms

  • Aquaporins / genetics
  • Aquaporins / metabolism*
  • Biological Transport
  • Cell Membrane / metabolism
  • Gene Expression
  • Gene Expression Regulation, Plant*
  • Hordeum / cytology
  • Hordeum / genetics*
  • Hordeum / metabolism
  • Hydrogen Peroxide / metabolism*
  • Mutagenesis, Site-Directed
  • Mutation
  • Organ Specificity
  • Oryza / cytology
  • Oryza / genetics*
  • Oryza / metabolism
  • Plant Epidermis / genetics
  • Plant Epidermis / metabolism
  • Plant Leaves / genetics
  • Plant Leaves / metabolism
  • Plant Proteins / genetics
  • Plant Proteins / metabolism
  • Plant Roots / genetics
  • Plant Roots / metabolism
  • Saccharomyces cerevisiae / cytology
  • Saccharomyces cerevisiae / genetics
  • Saccharomyces cerevisiae / metabolism
  • Serine / metabolism

Substances

  • Aquaporins
  • Plant Proteins
  • Serine
  • Hydrogen Peroxide