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Title: Identification of a plastidial phenylalanine exporter that influences flux distribution through the phenylalanine biosynthetic network

Abstract

In addition to proteins, L-phenylalanine is a versatile precursor for thousands of plant metabolites. Production of phenylalanine-derived compounds is a complex multi-compartmental process using phenylalanine synthesized predominantly in plastids as precursor. The transporter(s) exporting phenylalanine from plastids, however, remains unknown. Here, a gene encoding a Petunia hybrida plastidial cationic amino-acid transporter (PhpCAT) functioning in plastidial phenylalanine export is identified based on homology to an Escherichia coli phenylalanine transporter and co-expression with phenylalanine metabolic genes. Radiolabel transport assays show that PhpCAT exports all three aromatic amino acids. PhpCAT downregulation and overexpression result in decreased and increased levels, respectively, of phenylalanine-derived volatiles, as well as phenylalanine, tyrosine and their biosynthetic intermediates. Metabolic flux analysis reveals that flux through the plastidial phenylalanine biosynthetic pathway is reduced in PhpCAT RNAi lines, suggesting that the rate of phenylalanine export from plastids contributes to regulating flux through the aromatic amino-acid network.

Authors:
 [1];  [2];  [3];  [1];  [3];  [1];  [4];  [1];  [1];  [5];  [5];  [2];  [6];  [3]
  1. Purdue Univ., West Lafayette, IN (United States). Dept. of Biochemistry
  2. Purdue Univ., West Lafayette, IN (United States). Dept. of Horticulture and Landscape Architecture
  3. Purdue Univ., West Lafayette, IN (United States). Dept. of Biochemistry. Dept. of Horticulture and Landscape Architecture
  4. Purdue Univ., West Lafayette, IN (United States). School of Chemical Engineering
  5. Purdue Univ., West Lafayette, IN (United States). Bioinformatics Core
  6. Purdue Univ., West Lafayette, IN (United States). Dept. of Biochemistry. School of Chemical Engineering
Publication Date:
Research Org.:
Purdue Univ., West Lafayette, IN (United States)
Sponsoring Org.:
USDOE Office of Science (SC), Biological and Environmental Research (BER) (SC-23); Gordon and Betty Moore Foundation (United States)
OSTI Identifier:
1455141
Grant/Contract Number:  
SC0008628; GBMF2550.02
Resource Type:
Accepted Manuscript
Journal Name:
Nature Communications
Additional Journal Information:
Journal Volume: 6; Journal ID: ISSN 2041-1723
Publisher:
Nature Publishing Group
Country of Publication:
United States
Language:
English
Subject:
59 BASIC BIOLOGICAL SCIENCES

Citation Formats

Widhalm, Joshua R., Gutensohn, Michael, Yoo, Heejin, Adebesin, Funmilayo, Qian, Yichun, Guo, Longyun, Jaini, Rohit, Lynch, Joseph H., McCoy, Rachel M., Shreve, Jacob T., Thimmapuram, Jyothi, Rhodes, David, Morgan, John A., and Dudareva, Natalia. Identification of a plastidial phenylalanine exporter that influences flux distribution through the phenylalanine biosynthetic network. United States: N. p., 2015. Web. doi:10.1038/ncomms9142.
Widhalm, Joshua R., Gutensohn, Michael, Yoo, Heejin, Adebesin, Funmilayo, Qian, Yichun, Guo, Longyun, Jaini, Rohit, Lynch, Joseph H., McCoy, Rachel M., Shreve, Jacob T., Thimmapuram, Jyothi, Rhodes, David, Morgan, John A., & Dudareva, Natalia. Identification of a plastidial phenylalanine exporter that influences flux distribution through the phenylalanine biosynthetic network. United States. doi:10.1038/ncomms9142.
Widhalm, Joshua R., Gutensohn, Michael, Yoo, Heejin, Adebesin, Funmilayo, Qian, Yichun, Guo, Longyun, Jaini, Rohit, Lynch, Joseph H., McCoy, Rachel M., Shreve, Jacob T., Thimmapuram, Jyothi, Rhodes, David, Morgan, John A., and Dudareva, Natalia. Thu . "Identification of a plastidial phenylalanine exporter that influences flux distribution through the phenylalanine biosynthetic network". United States. doi:10.1038/ncomms9142. https://www.osti.gov/servlets/purl/1455141.
@article{osti_1455141,
title = {Identification of a plastidial phenylalanine exporter that influences flux distribution through the phenylalanine biosynthetic network},
author = {Widhalm, Joshua R. and Gutensohn, Michael and Yoo, Heejin and Adebesin, Funmilayo and Qian, Yichun and Guo, Longyun and Jaini, Rohit and Lynch, Joseph H. and McCoy, Rachel M. and Shreve, Jacob T. and Thimmapuram, Jyothi and Rhodes, David and Morgan, John A. and Dudareva, Natalia},
abstractNote = {In addition to proteins, L-phenylalanine is a versatile precursor for thousands of plant metabolites. Production of phenylalanine-derived compounds is a complex multi-compartmental process using phenylalanine synthesized predominantly in plastids as precursor. The transporter(s) exporting phenylalanine from plastids, however, remains unknown. Here, a gene encoding a Petunia hybrida plastidial cationic amino-acid transporter (PhpCAT) functioning in plastidial phenylalanine export is identified based on homology to an Escherichia coli phenylalanine transporter and co-expression with phenylalanine metabolic genes. Radiolabel transport assays show that PhpCAT exports all three aromatic amino acids. PhpCAT downregulation and overexpression result in decreased and increased levels, respectively, of phenylalanine-derived volatiles, as well as phenylalanine, tyrosine and their biosynthetic intermediates. Metabolic flux analysis reveals that flux through the plastidial phenylalanine biosynthetic pathway is reduced in PhpCAT RNAi lines, suggesting that the rate of phenylalanine export from plastids contributes to regulating flux through the aromatic amino-acid network.},
doi = {10.1038/ncomms9142},
journal = {Nature Communications},
number = ,
volume = 6,
place = {United States},
year = {2015},
month = {9}
}

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