AtRAB-H1b and AtRAB-H1c GTPases, homologues of the yeast Ypt6, target reporter proteins to the Golgi when expressed in Nicotiana tabacum and Arabidopsis thaliana

Abstract

Ypt/Rab GTPases act as key regulators of intracellular traffic through the conformational differences exhibited by their GTP or GDP-bound forms. In this paper, two Arabidopsis Ypt6 homologues, AtRAB-H1b and AtRAB-H1 c were characterized and compared. Using a live cell imaging approach, it is shown that yellow fluorescent protein-fusions (YFP) of AtRAB-H1b and AtRAB-H1c locate to the Golgi and to the cytosol in both Nicotiana tabacum and in Arabidopsis thaliana. In addition, YFP-AtRAB-H1b targets an as yet unknown compartment not labelled by YFP-AtRAB-H1c or Golgi markers. It is also shown that the subcellular location of YFP-AtRAB-H1b and YFP-AtRAB-H1c is affected by the state of GTP-binding and that expression of a GTP-deficient mutant results in increased apoplastic fluorescence of a secretory form of YFP.