ABCB19/PGP19 stabilises PIN1 in membrane microdomains in Arabidopsis

Boosaree Titapiwatanakun, Joshua J. Blakeslee, Anindita Bandyopadhyay, Haibing Yang, Jozef Mravec, Michael Sauer, Yan Cheng, Jiri Adamec, Akitomo Nagashima, Markus Geisler, Tatsuya Sakai, Jiri Friml, Wendy Ann Peer, Angus S. Murphy

Research output: Contribution to journalArticle

165 Scopus citations

Abstract

Auxin transport is mediated at the cellular level by three independent mechanisms that are characterised by the PIN-formed (PIN), P-glycoprotein (ABCB/PGP) and AUX/LAX transport proteins. The PIN and ABCB transport proteins, best represented by PIN1 and ABCB19 (PGP19), have been shown to coordinately regulate auxin efflux. When PIN1 and ABCB19 coincide on the plasma membrane, their interaction enhances the rate and specificity of auxin efflux and the dynamic cycling of PIN1 is reduced. However, ABCB19 function is not regulated by the dynamic cellular trafficking mechanisms that regulate PIN1 in apical tissues, as localisation of ABCB19 on the plasma membrane was not inhibited by short-term treatments with latrunculin B, oryzalin, brefeldin A (BFA) or wortmannin - all of which have been shown to alter PIN1 and/or PIN2 plasma membrane localisation. When taken up by endocytosis, the styryl dye FM4-64 labels diffuse rather than punctuate intracellular bodies in abcb19 (pgp19), and some aggregations of PIN1 induced by short-term BFA treatment did not disperse after BFA washout in abcb19. Although the subcellular localisations of ABCB19 and PIN1 in the reciprocal mutant backgrounds were like those in wild type, PIN1 plasma membrane localisation in abcb19 roots was more easily perturbed by the detergent Triton X-100, but not other non-ionic detergents. ABCB19 is stably associated with sterol/sphingolipid-enriched membrane fractions containing BIG/TIR3 and partitions into Triton X-100 detergent-resistant membrane (DRM) fractions. In the wild type, PIN1 was also present in DRMs, but was less abundant in abcb19 DRMs. These observations suggested a rationale for the observed lack of auxin transport activity when PIN1 is expressed in a non-plant heterologous system. PIN1 was therefore expressed in Schizosaccharomyces pombe, which has plant-like sterol-enriched microdomains, and catalysed auxin transport in these cells. These data suggest that ABCB19 stabilises PIN1 localisation at the plasma membrane in discrete cellular subdomains where PIN1 and ABCB19 expression overlaps.

Original languageEnglish (US)
Pages (from-to)27-44
Number of pages18
JournalPlant Journal
Volume57
Issue number1
DOIs
StatePublished - Jan 2009

Keywords

  • Auxin transport
  • Detergent-resistant membrane
  • Lipid raft
  • Multi-drug resistance protein
  • P-glycoprotein
  • PIN1
  • Trafficking

ASJC Scopus subject areas

  • Genetics
  • Plant Science
  • Cell Biology

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    Titapiwatanakun, B., Blakeslee, J. J., Bandyopadhyay, A., Yang, H., Mravec, J., Sauer, M., Cheng, Y., Adamec, J., Nagashima, A., Geisler, M., Sakai, T., Friml, J., Peer, W. A., & Murphy, A. S. (2009). ABCB19/PGP19 stabilises PIN1 in membrane microdomains in Arabidopsis. Plant Journal, 57(1), 27-44. https://doi.org/10.1111/j.1365-313X.2008.03668.x