Deletion analysis of AGD1 reveals domains crucial for plasma membrane recruitment and function in root hair polarity

J Cell Sci. 2018 Jan 29;131(2):jcs203828. doi: 10.1242/jcs.203828.

Abstract

AGD1, a plant ACAP-type ADP-ribosylation factor-GTPase activating protein (ARF-GAP), functions in specifying root hair polarity in Arabidopsis thaliana To better understand how AGD1 modulates root hair growth, we generated full-length and domain-deleted AGD1-green fluorescent protein (GFP) constructs, and followed their localization during root hair development. AGD1-GFP localized to the cytoplasm and was recruited to specific regions of the root hair plasma membrane (PM). Distinct PM AGD1-GFP signal was first detected along the site of root hair bulge formation. The construct continued to mark the PM at the root hair apical dome, but only during periods of reduced growth. During rapid tip growth, AGD1-GFP labeled the PM of the lateral flanks and dissipated from the apical-most PM. Deletion analysis and a single domain GFP fusion revealed that the pleckstrin homology (PH) domain is the minimal unit required for recruitment of AGD1 to the PM. Our results indicate that differential recruitment of AGD1 to specific PM domains is an essential component of the membrane trafficking machinery that facilitates root hair developmental phase transitions and responses to changes in the root microenvironment.

Keywords: ADP-ribosylation factor-GTPase activating protein; Arabidopsis thaliana; Plasma membrane; Polarity; Root hairs; Tip growth.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Arabidopsis Proteins / chemistry*
  • Arabidopsis Proteins / metabolism*
  • Cell Membrane / metabolism*
  • Cytoplasm / metabolism
  • GTPase-Activating Proteins / chemistry*
  • GTPase-Activating Proteins / metabolism*
  • Green Fluorescent Proteins / metabolism
  • Mutation / genetics
  • Phosphatidylinositol Phosphates / metabolism
  • Plant Roots / growth & development
  • Plant Roots / metabolism*
  • Protein Domains
  • Recombinant Fusion Proteins / metabolism
  • Sequence Deletion*
  • Structure-Activity Relationship

Substances

  • AGD1 protein, Arabidopsis
  • Arabidopsis Proteins
  • GTPase-Activating Proteins
  • Phosphatidylinositol Phosphates
  • Recombinant Fusion Proteins
  • phosphatidylinositol 4-phosphate
  • Green Fluorescent Proteins