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Transport activity of rice sucrose transporters OsSUT1 and OsSUT5.

ABSTRACT: Expression in Xenopus oocytes and electrophysiology was used to test for transport activity of the five sucrose transporter (SUT) homologs from rice. Expression of OsSUT1 and OsSUT5 resulted in sucrose-dependent currents that were analyzed by two-electrode voltage clamping. We examined the transport kinetics, substrate specificity and pH dependence of sucrose transport and K(0.5) for sucrose. OsSUT1 showed similar features to those of other type II SUTs from monocots examined previously, with a K(0.5) value of 7.50 mM at pH 5.6. In contrast, OsSUT5 had a higher substrate affinity (K(0.5) = 2.32 mM at pH 5.6), less substrate specificity and less pH dependence compared with all type II SUTs tested to date. Regulation of the rice SUTs, as well as ZmSUT1 from maize and HvSUT1 from barley, by reduced (GSH) and oxidized (GSSG) forms of glutathione was tested. GSSG and GSH were found to have no significant effect on the activity of sucrose transporters when expressed in Xenopus oocytes. In conclusion, differences in transport activity between OsSUT1 and OsSUT5 indicate that type II SUTs have a range of transport activities that are tuned to their function in the plant.

SUBMITTER: Sun Y

PROVIDER: S-EPMC2807175 | biostudies-literature | 2010 Jan

REPOSITORIES: biostudies-literature

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Transport activity of rice sucrose transporters OsSUT1 and OsSUT5.

Sun Ye Y Reinders Anke A LaFleur Kathryn R KR Mori Toko T Ward John M JM

Plant & cell physiology 20091203 1

Expression in Xenopus oocytes and electrophysiology was used to test for transport activity of the five sucrose transporter (SUT) homologs from rice. Expression of OsSUT1 and OsSUT5 resulted in sucrose-dependent currents that were analyzed by two-electrode voltage clamping. We examined the transport kinetics, substrate specificity and pH dependence of sucrose transport and K(0.5) for sucrose. OsSUT1 showed similar features to those of other type II SUTs from monocots examined previously, with a ...[more]

PMID: 19965875

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Project description:The best characterized function of sucrose transporters of the SUC family in plants is the uptake of sucrose into the phloem for long-distance transport of photoassimilates. This important step is usually performed by one specific SUC in every species. However, plants possess small families of several different SUCs which are less well understood. Here, we report on the characterization of AtSUC6 and AtSUC7, two members of the SUC family in Arabidopsis thaliana. Heterologous expression in yeast (Saccharomyces cerevisiae) revealed that AtSUC6Col-0 is a high-affinity H+-symporter that mediates the uptake of sucrose and maltose across the plasma membrane at exceptionally low pH values. Reporter gene analyses revealed a strong expression of AtSUC6Col-0 in reproductive tissues, where the protein product might contribute to sugar uptake into pollen tubes and synergid cells. A knockout of AtSUC6 did not interfere with vegetative development or reproduction, which points toward physiological redundancy of AtSUC6Col-0 with other sugar transporters. Reporter gene analyses showed that AtSUC7Col-0 is expressed in roots and pollen tubes and that this sink specific expression of AtSUC7Col-0 is regulated by intragenic regions. Transport activity of AtSUC7Col-0 could not be analyzed in baker's yeast or Xenopus oocytes because the protein was not correctly targeted to the plasma membrane in both heterologous expression systems. Therefore, a novel approach to analyze sucrose transporters in planta was developed. Plasma membrane localized SUCs including AtSUC6Col-0 and also sucrose specific SWEETs were able to mediate transport of the fluorescent sucrose analog esculin in transformed mesophyll protoplasts. In contrast, AtSUC7Col-0 is not able to mediate esculin transport across the plasma membrane which implicates that AtSUC7Col-0 might be a non-functional pseudogene. The novel protoplast assay provides a useful tool for the quick and quantitative analysis of sucrose transporters in an in planta expression system.

Dataset Information

Transport activity of rice sucrose transporters OsSUT1 and OsSUT5.

Publications

Transport activity of rice sucrose transporters OsSUT1 and OsSUT5.

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