Interactions between co-expressed Arabidopsis sucrose transporters in the split-ubiquitin system

Waltraud X. Schulze, Anke Reinders, John Ward, Sylvie Lalonde, Wolf B. Frommer

Research output: Contribution to journalArticle

55 Citations (Scopus)

Abstract

Background: The Arabidopsis genome contains nine sucrose transporter paralogs falling into three clades: SUT1-like, SUT2 and SUT4. The carriers differ in their kinetic properties. Many transport proteins are known to exist as oligomers. The yeast-based split ubiquitin system can be used to analyze the ability of membrane proteins to interact. Results: Promoter-GUS fusions were used to analyze the cellular expression of the three transporter genes in transgenic Arabidopsis plants. All three fusion genes are co-expressed in companion cells. Protein-protein interactions between Arabidopsis sucrose transporters were tested using the split ubiquitin system. Three paralogous sucrose transporters are capable of interacting as either homo- or heteromers. The interactions are specific, since a potassium channel and a glucose transporter did not show interaction with sucrose transporters. Also the biosynthetic and metabolizing enzymes, sucrose phosphate phosphatase and sucrose synthase, which were found to be at least in part bound to the plasma membrane, did not specifically interact with sucrose transporters. Conclusions: The split-ubiquitin system provides a powerful tool to detect potential interactions between plant membrane proteins by heterologous expression in yeast, and can be used to screen for interactions with membrane proteins as baits. Like other membrane proteins, the Arabidopsis sucrose transporters are able to form oligomers. The biochemical approaches are required to confirm the in planta interaction.

Original languageEnglish (US)
Article number1
Pages (from-to)1-10
Number of pages10
JournalBMC Biochemistry
Volume4
Issue number1
DOIs
StatePublished - Mar 18 2003

Fingerprint

Ubiquitin
Arabidopsis
Sucrose
Membrane Proteins
Genes
Oligomers
Yeast
Fusion reactions
Yeasts
Facilitative Glucose Transport Proteins
Potassium Channels
Phosphatases
Sugar (sucrose)
Plant Proteins
Cell membranes
Gene Fusion
Genetically Modified Plants
Carrier Proteins
Phosphates
Proteins

Cite this

Interactions between co-expressed Arabidopsis sucrose transporters in the split-ubiquitin system. / Schulze, Waltraud X.; Reinders, Anke; Ward, John; Lalonde, Sylvie; Frommer, Wolf B.

In: BMC Biochemistry, Vol. 4, No. 1, 1, 18.03.2003, p. 1-10.

Research output: Contribution to journalArticle

Schulze, Waltraud X. ; Reinders, Anke ; Ward, John ; Lalonde, Sylvie ; Frommer, Wolf B. / Interactions between co-expressed Arabidopsis sucrose transporters in the split-ubiquitin system. In: BMC Biochemistry. 2003 ; Vol. 4, No. 1. pp. 1-10.
@article{c05583b72feb476094425a2bf84bf6c7,
title = "Interactions between co-expressed Arabidopsis sucrose transporters in the split-ubiquitin system",
abstract = "Background: The Arabidopsis genome contains nine sucrose transporter paralogs falling into three clades: SUT1-like, SUT2 and SUT4. The carriers differ in their kinetic properties. Many transport proteins are known to exist as oligomers. The yeast-based split ubiquitin system can be used to analyze the ability of membrane proteins to interact. Results: Promoter-GUS fusions were used to analyze the cellular expression of the three transporter genes in transgenic Arabidopsis plants. All three fusion genes are co-expressed in companion cells. Protein-protein interactions between Arabidopsis sucrose transporters were tested using the split ubiquitin system. Three paralogous sucrose transporters are capable of interacting as either homo- or heteromers. The interactions are specific, since a potassium channel and a glucose transporter did not show interaction with sucrose transporters. Also the biosynthetic and metabolizing enzymes, sucrose phosphate phosphatase and sucrose synthase, which were found to be at least in part bound to the plasma membrane, did not specifically interact with sucrose transporters. Conclusions: The split-ubiquitin system provides a powerful tool to detect potential interactions between plant membrane proteins by heterologous expression in yeast, and can be used to screen for interactions with membrane proteins as baits. Like other membrane proteins, the Arabidopsis sucrose transporters are able to form oligomers. The biochemical approaches are required to confirm the in planta interaction.",
author = "Schulze, {Waltraud X.} and Anke Reinders and John Ward and Sylvie Lalonde and Frommer, {Wolf B.}",
year = "2003",
month = "3",
day = "18",
doi = "10.1186/1471-2091-4-3",
language = "English (US)",
volume = "4",
pages = "1--10",
journal = "BMC Biochemistry",
issn = "1471-2091",
publisher = "BioMed Central",
number = "1",

}

TY - JOUR

T1 - Interactions between co-expressed Arabidopsis sucrose transporters in the split-ubiquitin system

AU - Schulze, Waltraud X.

AU - Reinders, Anke

AU - Ward, John

AU - Lalonde, Sylvie

AU - Frommer, Wolf B.

PY - 2003/3/18

Y1 - 2003/3/18

N2 - Background: The Arabidopsis genome contains nine sucrose transporter paralogs falling into three clades: SUT1-like, SUT2 and SUT4. The carriers differ in their kinetic properties. Many transport proteins are known to exist as oligomers. The yeast-based split ubiquitin system can be used to analyze the ability of membrane proteins to interact. Results: Promoter-GUS fusions were used to analyze the cellular expression of the three transporter genes in transgenic Arabidopsis plants. All three fusion genes are co-expressed in companion cells. Protein-protein interactions between Arabidopsis sucrose transporters were tested using the split ubiquitin system. Three paralogous sucrose transporters are capable of interacting as either homo- or heteromers. The interactions are specific, since a potassium channel and a glucose transporter did not show interaction with sucrose transporters. Also the biosynthetic and metabolizing enzymes, sucrose phosphate phosphatase and sucrose synthase, which were found to be at least in part bound to the plasma membrane, did not specifically interact with sucrose transporters. Conclusions: The split-ubiquitin system provides a powerful tool to detect potential interactions between plant membrane proteins by heterologous expression in yeast, and can be used to screen for interactions with membrane proteins as baits. Like other membrane proteins, the Arabidopsis sucrose transporters are able to form oligomers. The biochemical approaches are required to confirm the in planta interaction.

AB - Background: The Arabidopsis genome contains nine sucrose transporter paralogs falling into three clades: SUT1-like, SUT2 and SUT4. The carriers differ in their kinetic properties. Many transport proteins are known to exist as oligomers. The yeast-based split ubiquitin system can be used to analyze the ability of membrane proteins to interact. Results: Promoter-GUS fusions were used to analyze the cellular expression of the three transporter genes in transgenic Arabidopsis plants. All three fusion genes are co-expressed in companion cells. Protein-protein interactions between Arabidopsis sucrose transporters were tested using the split ubiquitin system. Three paralogous sucrose transporters are capable of interacting as either homo- or heteromers. The interactions are specific, since a potassium channel and a glucose transporter did not show interaction with sucrose transporters. Also the biosynthetic and metabolizing enzymes, sucrose phosphate phosphatase and sucrose synthase, which were found to be at least in part bound to the plasma membrane, did not specifically interact with sucrose transporters. Conclusions: The split-ubiquitin system provides a powerful tool to detect potential interactions between plant membrane proteins by heterologous expression in yeast, and can be used to screen for interactions with membrane proteins as baits. Like other membrane proteins, the Arabidopsis sucrose transporters are able to form oligomers. The biochemical approaches are required to confirm the in planta interaction.

UR - http://www.scopus.com/inward/record.url?scp=2942683234&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=2942683234&partnerID=8YFLogxK

U2 - 10.1186/1471-2091-4-3

DO - 10.1186/1471-2091-4-3

M3 - Article

C2 - 12689351

AN - SCOPUS:0347386495

VL - 4

SP - 1

EP - 10

JO - BMC Biochemistry

JF - BMC Biochemistry

SN - 1471-2091

IS - 1

M1 - 1

ER -