a framework for lateral membrane trafficking and polar tethering of the pen3 atp binding cassette transporter

Nature 433: 39-44 Pubmed: Author and Title CrossRef: Author and Title Google Scholar: Author Only Title Only Author and Title Boutté Y, Crosnier MT, Carraro N, Traas J, Satiat-Jeunemaitr

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Running title: PEN3 outer lateral domain trafficking and polarity

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TRANSPORTER1 (IRT1; Barberon et al., 2014), the boron exporter BOR4

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To address a potential contribution of endocytosis, we applied the endocytosis

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To more directly monitor PEN3 endocytosis and recycling, we generated

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PEN3 endocytic and secretory trafficking via the trans-Golgi network

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to the TGN in vivo, we generated transgenic plants co-expressing PEN3-GFP

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showed that 48% of intracellular PEN3-GFP co-localized with SYP61-CFP

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organization Consistent with previous observations (Łangowski et al., 2010),

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In comparison, the actin depolymerizing drug latrunculin B (LatB) induced

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To identify the actin isoform involved, we examined PEN3-GFP distribution in

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methanesulfonate (EMS)-mutagenized population We identified three mutant

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Similarly, act7-7 revealed a PEN3-GFP mis-localization phenotype

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(Fig 6, Q and R) The decrease of red PEN3-mEos2 signal in act7-6

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8A; Supplemental Fig S4A), while in the exo84b-1 mutant FRAP of

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the cell The polar index of the wild type (9.95 ± 1.22) proved clearly and

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for ACT7 function in trafficking and tethering of outer lateral membrane

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MATERIALS AND METHODS

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used at 10 μM for 2.5 h from a 100 mM stock solution in DMSO In control

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seedlings were either treated with BFA, or with 1 h CHX pre-treatment

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bleaching Within these cells, 30 bleaching iterations were performed at five to

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compartments was measured at indicated time points by Fiji software

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Transmission electron microscopy

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ligation at the SmaI site The coding sequence of mEos2 (McKinney et al.,

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the H2B fused to mCherry, the region of Venus in

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F1 indicating allelelism The coding sequence of ACT7 was amplified and

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Statistical differences of BFA body size (J) and fluorescence intensity per BFA

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Figure 3 Cytoplasmic PEN3 preferentially co-localizes with trans-Golgi

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FM4-64 (FM, magenta) and two washes E to H, Cells expressing PEN3-GFP

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independent roots observed with similar results M to P, PEN3-mEos2

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FM4-64 (FM, magenta) for 5 min in (F) wild type (WT) and (G) exo84b-1 (H),

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mean intensity of outer lateral PM domain to obtain the relative fluorescence

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