Functional identification of Arabidopsis ATSIP2 (At3g57520) as an alkaline α-galactosidase with a substrate specificity for raffinose and an apparent sink-specific expression pattern

Plant Cell Physiol. 2010 Oct;51(10):1815-9. doi: 10.1093/pcp/pcq127. Epub 2010 Aug 24.

Abstract

Arabidopsis ATSIP2 has recently been suggested to be a raffinose synthase gene. However, it has high amino acid identity to functionally characterized alkaline α-galactosidases from Cucumis melo and Zea mays. Using the Sf9 insect cell expression system, we demonstrate that recombinant ATSIP2 is a genuine alkaline α-galactosidase with a distinct substrate specificity for raffinose, and not a raffinose synthase. A β-glucuronidase reporter construct using the ATSIP2 promoter shows that ATSIP2 is strongly expressed in sink tissues of Arabidopsis, i.e. sink leaves and non-xylem parts of the root stele, suggesting a physiological function in raffinose phloem unloading.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Animals
  • Arabidopsis / genetics*
  • Arabidopsis / metabolism
  • Arabidopsis Proteins / genetics
  • Arabidopsis Proteins / metabolism*
  • Cell Line
  • Gene Expression Regulation, Plant
  • Insecta / cytology
  • Phloem / metabolism*
  • Promoter Regions, Genetic
  • RNA, Plant / genetics
  • Raffinose / metabolism*
  • Recombinant Proteins / genetics
  • Recombinant Proteins / metabolism
  • Substrate Specificity
  • alpha-Galactosidase / genetics
  • alpha-Galactosidase / metabolism*

Substances

  • Arabidopsis Proteins
  • RNA, Plant
  • Recombinant Proteins
  • alpha-Galactosidase
  • Raffinose