Identification and characterization of the Na+/H+ antiporter NhaS3 from the thylakoid membrane of Synechocystis sp. PCC 6803

Kenta Tsunekawa, Toshiaki Shijuku, Mitsuo Hayashimoto, Yoichi Kojima, Kiyoshi Onai, Megumi Morishita, Masahiro Ishiura, Teruo Kuroda, Tatsunosuke Nakamura, Hiroshi Kobayashi, Mayuko Sato, Kiminori Toyooka, Ken Matsuoka, Tatsuo Omata, Nobuyuki Uozumi

Research output: Contribution to journalArticle

36 Citations (Scopus)

Abstract

Na+/H+ antiporters influence proton or sodium motive force across the membrane. Synechocystis sp. PCC 6803 has six genes encoding Na+/H+ antiporters, nhaS1-5 and sll0556. In this study, the function of NhaS3 was examined. NhaS3 was essential for growth of Synechocystis, and loss of nhaS3 was not complemented by expression of the Escherichia coli Na+/ H+ antiporter NhaA. Membrane fractionation followed by immunoblotting as well as immunogold labeling revealed that NhaS3 was localized in the thylakoid membrane of Synechocystis. NhaS3 was shown to be functional over apHrange from pH 6.5 to 9.0 when expressed in E. coli. A reduction in the copy number of nhaS3 in the Synechocystis genome rendered the cells more sensitive to high Na+ concentrations. NhaS3 had no K+/H+ exchange activity itself but enhanced K+ uptake from the medium when expressed in an E. coli potassium uptake mutant. Expression of nhaS3 increased after shifting from low CO2 to high CO2 conditions. Expression of nhaS3 was also found to be controlled by the circadian rhythm. Gene expression peaked at the beginning of subjective night. This coincided with the time of the lowest rate of CO2 consumption caused by the ceasing of O2-evolving photosynthesis. This is the first report of a Na+/H+ antiporter localized in thylakoid membrane. Our results suggested a role of NhaS3 in the maintenance of ion homeostasis of H+, Na+, and K+ in supporting the conversion of photosynthetic products and in the supply of energy in the dark.

Original languageEnglish
Pages (from-to)16513-16521
Number of pages9
JournalJournal of Biological Chemistry
Volume284
Issue number24
DOIs
Publication statusPublished - Jun 12 2009

Fingerprint

Synechocystis
Sodium-Hydrogen Antiporter
Thylakoids
Escherichia coli
Membranes
Gene encoding
Photosynthesis
Fractionation
Circadian Rhythm
Immunoblotting
Gene expression
Labeling
Protons
Potassium
Homeostasis
Genes
Sodium
Maintenance
Genome
Ions

All Science Journal Classification (ASJC) codes

  • Biochemistry
  • Molecular Biology
  • Cell Biology

Cite this

Tsunekawa, K., Shijuku, T., Hayashimoto, M., Kojima, Y., Onai, K., Morishita, M., ... Uozumi, N. (2009). Identification and characterization of the Na+/H+ antiporter NhaS3 from the thylakoid membrane of Synechocystis sp. PCC 6803. Journal of Biological Chemistry, 284(24), 16513-16521. https://doi.org/10.1074/jbc.M109.001875

Identification and characterization of the Na+/H+ antiporter NhaS3 from the thylakoid membrane of Synechocystis sp. PCC 6803. / Tsunekawa, Kenta; Shijuku, Toshiaki; Hayashimoto, Mitsuo; Kojima, Yoichi; Onai, Kiyoshi; Morishita, Megumi; Ishiura, Masahiro; Kuroda, Teruo; Nakamura, Tatsunosuke; Kobayashi, Hiroshi; Sato, Mayuko; Toyooka, Kiminori; Matsuoka, Ken; Omata, Tatsuo; Uozumi, Nobuyuki.

In: Journal of Biological Chemistry, Vol. 284, No. 24, 12.06.2009, p. 16513-16521.

Research output: Contribution to journalArticle

Tsunekawa, K, Shijuku, T, Hayashimoto, M, Kojima, Y, Onai, K, Morishita, M, Ishiura, M, Kuroda, T, Nakamura, T, Kobayashi, H, Sato, M, Toyooka, K, Matsuoka, K, Omata, T & Uozumi, N 2009, 'Identification and characterization of the Na+/H+ antiporter NhaS3 from the thylakoid membrane of Synechocystis sp. PCC 6803', Journal of Biological Chemistry, vol. 284, no. 24, pp. 16513-16521. https://doi.org/10.1074/jbc.M109.001875
Tsunekawa, Kenta ; Shijuku, Toshiaki ; Hayashimoto, Mitsuo ; Kojima, Yoichi ; Onai, Kiyoshi ; Morishita, Megumi ; Ishiura, Masahiro ; Kuroda, Teruo ; Nakamura, Tatsunosuke ; Kobayashi, Hiroshi ; Sato, Mayuko ; Toyooka, Kiminori ; Matsuoka, Ken ; Omata, Tatsuo ; Uozumi, Nobuyuki. / Identification and characterization of the Na+/H+ antiporter NhaS3 from the thylakoid membrane of Synechocystis sp. PCC 6803. In: Journal of Biological Chemistry. 2009 ; Vol. 284, No. 24. pp. 16513-16521.
@article{3b7de61425c0432e87a4c1093f24f9ad,
title = "Identification and characterization of the Na+/H+ antiporter NhaS3 from the thylakoid membrane of Synechocystis sp. PCC 6803",
abstract = "Na+/H+ antiporters influence proton or sodium motive force across the membrane. Synechocystis sp. PCC 6803 has six genes encoding Na+/H+ antiporters, nhaS1-5 and sll0556. In this study, the function of NhaS3 was examined. NhaS3 was essential for growth of Synechocystis, and loss of nhaS3 was not complemented by expression of the Escherichia coli Na+/ H+ antiporter NhaA. Membrane fractionation followed by immunoblotting as well as immunogold labeling revealed that NhaS3 was localized in the thylakoid membrane of Synechocystis. NhaS3 was shown to be functional over apHrange from pH 6.5 to 9.0 when expressed in E. coli. A reduction in the copy number of nhaS3 in the Synechocystis genome rendered the cells more sensitive to high Na+ concentrations. NhaS3 had no K+/H+ exchange activity itself but enhanced K+ uptake from the medium when expressed in an E. coli potassium uptake mutant. Expression of nhaS3 increased after shifting from low CO2 to high CO2 conditions. Expression of nhaS3 was also found to be controlled by the circadian rhythm. Gene expression peaked at the beginning of subjective night. This coincided with the time of the lowest rate of CO2 consumption caused by the ceasing of O2-evolving photosynthesis. This is the first report of a Na+/H+ antiporter localized in thylakoid membrane. Our results suggested a role of NhaS3 in the maintenance of ion homeostasis of H+, Na+, and K+ in supporting the conversion of photosynthetic products and in the supply of energy in the dark.",
author = "Kenta Tsunekawa and Toshiaki Shijuku and Mitsuo Hayashimoto and Yoichi Kojima and Kiyoshi Onai and Megumi Morishita and Masahiro Ishiura and Teruo Kuroda and Tatsunosuke Nakamura and Hiroshi Kobayashi and Mayuko Sato and Kiminori Toyooka and Ken Matsuoka and Tatsuo Omata and Nobuyuki Uozumi",
year = "2009",
month = "6",
day = "12",
doi = "10.1074/jbc.M109.001875",
language = "English",
volume = "284",
pages = "16513--16521",
journal = "Journal of Biological Chemistry",
issn = "0021-9258",
publisher = "American Society for Biochemistry and Molecular Biology Inc.",
number = "24",

}

TY - JOUR

T1 - Identification and characterization of the Na+/H+ antiporter NhaS3 from the thylakoid membrane of Synechocystis sp. PCC 6803

AU - Tsunekawa, Kenta

AU - Shijuku, Toshiaki

AU - Hayashimoto, Mitsuo

AU - Kojima, Yoichi

AU - Onai, Kiyoshi

AU - Morishita, Megumi

AU - Ishiura, Masahiro

AU - Kuroda, Teruo

AU - Nakamura, Tatsunosuke

AU - Kobayashi, Hiroshi

AU - Sato, Mayuko

AU - Toyooka, Kiminori

AU - Matsuoka, Ken

AU - Omata, Tatsuo

AU - Uozumi, Nobuyuki

PY - 2009/6/12

Y1 - 2009/6/12

N2 - Na+/H+ antiporters influence proton or sodium motive force across the membrane. Synechocystis sp. PCC 6803 has six genes encoding Na+/H+ antiporters, nhaS1-5 and sll0556. In this study, the function of NhaS3 was examined. NhaS3 was essential for growth of Synechocystis, and loss of nhaS3 was not complemented by expression of the Escherichia coli Na+/ H+ antiporter NhaA. Membrane fractionation followed by immunoblotting as well as immunogold labeling revealed that NhaS3 was localized in the thylakoid membrane of Synechocystis. NhaS3 was shown to be functional over apHrange from pH 6.5 to 9.0 when expressed in E. coli. A reduction in the copy number of nhaS3 in the Synechocystis genome rendered the cells more sensitive to high Na+ concentrations. NhaS3 had no K+/H+ exchange activity itself but enhanced K+ uptake from the medium when expressed in an E. coli potassium uptake mutant. Expression of nhaS3 increased after shifting from low CO2 to high CO2 conditions. Expression of nhaS3 was also found to be controlled by the circadian rhythm. Gene expression peaked at the beginning of subjective night. This coincided with the time of the lowest rate of CO2 consumption caused by the ceasing of O2-evolving photosynthesis. This is the first report of a Na+/H+ antiporter localized in thylakoid membrane. Our results suggested a role of NhaS3 in the maintenance of ion homeostasis of H+, Na+, and K+ in supporting the conversion of photosynthetic products and in the supply of energy in the dark.

AB - Na+/H+ antiporters influence proton or sodium motive force across the membrane. Synechocystis sp. PCC 6803 has six genes encoding Na+/H+ antiporters, nhaS1-5 and sll0556. In this study, the function of NhaS3 was examined. NhaS3 was essential for growth of Synechocystis, and loss of nhaS3 was not complemented by expression of the Escherichia coli Na+/ H+ antiporter NhaA. Membrane fractionation followed by immunoblotting as well as immunogold labeling revealed that NhaS3 was localized in the thylakoid membrane of Synechocystis. NhaS3 was shown to be functional over apHrange from pH 6.5 to 9.0 when expressed in E. coli. A reduction in the copy number of nhaS3 in the Synechocystis genome rendered the cells more sensitive to high Na+ concentrations. NhaS3 had no K+/H+ exchange activity itself but enhanced K+ uptake from the medium when expressed in an E. coli potassium uptake mutant. Expression of nhaS3 increased after shifting from low CO2 to high CO2 conditions. Expression of nhaS3 was also found to be controlled by the circadian rhythm. Gene expression peaked at the beginning of subjective night. This coincided with the time of the lowest rate of CO2 consumption caused by the ceasing of O2-evolving photosynthesis. This is the first report of a Na+/H+ antiporter localized in thylakoid membrane. Our results suggested a role of NhaS3 in the maintenance of ion homeostasis of H+, Na+, and K+ in supporting the conversion of photosynthetic products and in the supply of energy in the dark.

UR - http://www.scopus.com/inward/record.url?scp=67650221596&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=67650221596&partnerID=8YFLogxK

U2 - 10.1074/jbc.M109.001875

DO - 10.1074/jbc.M109.001875

M3 - Article

C2 - 19372598

AN - SCOPUS:67650221596

VL - 284

SP - 16513

EP - 16521

JO - Journal of Biological Chemistry

JF - Journal of Biological Chemistry

SN - 0021-9258

IS - 24

ER -