Functional characterization of the uracil transporter from honeybee pathogen Paenibacillus larvae

Stoffer-Bittner, Amanda J.; Alexander, Candace R; Dingman, Douglas W.; Mourad, George; Schultes, Neil P.

doi:10.1016/j.micpath.2018.08.059

Cited by 4 publications

(1 citation statement)

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“…To assess the thermodynamic aspect of the uptake of tZ, tZR, iP, and iPR in the BY-2 cells, we performed a series of accumulation assays in suspensions treated with 50 µM protonophore carbonyl cyanide 3-chlorophenylhydrazone (CCCP) in dimethyl sulfoxide (DMSO). CCCP uncouples electron transfer from oxidative phosphorylation (Cavari et al, 1967; Cunarro and Weiner, 1975; Heytler, 1963), thus inhibiting proton gradient-dependent transport processes (Alexander et al, 2018; Culos and Watanabe, 1983; Stoffer-Bittner et al, 2018). We performed the same assays using cells treated with the corresponding amount of DMSO alone (mock treatment) as control.…”

Section: Resultsmentioning

confidence: 99%

Comprehensive Model of Cell-to-Cell Cytokinin Transport Reveals A Specific Mode of Cytokinin Riboside Influx

Nedvěd,

Hudeček,

Klíma

et al. 2024

Preprint

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1AbstractRibosylated forms of plant hormones cytokinins (CKs) are the dominant CK species translocated at long distances. Their particular roles in plant physiology imply the existence of a yet uncharacterized CK riboside-specific membrane transport system. In this work, we report significant differences in the kinetics of the membrane transport of CK nucleobases and ribosides and the overall affinity of membrane-bound carriers towards the two CK forms. We show that CK ribosides can inhibit the uptake of CK nucleobases in tobacco Bright Yellow 2 cell suspensions but not vice versa, confirming the existence of a membrane transport system that strictly recognizes CK ribosides.We further characterize the membrane transport of CK nucleobases and ribosides mediated by AtENT3 (EQULIBRATIVE NUCLEOSIDE TRANSPORTER 3), showing its preference towardstrans-zeatin riboside (tZR) over isopentenyl adenosine (iPR). With the molecular docking and molecular dynamics, we assess the interactions among the side chain of tZR and AtENT3 residues Tyr61 and Asp129, which are conserved in all AtENTs but not in the ENTs from non-plant species. Lastly, we show thatatent3mutation affects shoot phenotype, demonstrating the impact of CK riboside membrane transport on shoot development.

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