An assembly factor promotes assembly of flavinated SDH1 into the succinate dehydrogenase complex

Katharina Belt, Olivier Van Aken, Monika Murcha, Andrew Millar, Shaobai Huang

Research output: Contribution to journalArticle

4 Citations (Scopus)

Abstract

Succinate dehydrogenase (Complex II; SDH) plays an important role in mitochondrial respiratory metabolism. The SDH complex consists of four core subunits and multiple cofactors, which must be assembled correctly to ensure enzyme function. To date, only an assembly factor (SDHAF2) required for FAD insertion into subunit SDH1 has been identified in plants. Here, we report the identification of Arabidopsis (Arabidopsis thaliana) At5g67490 as a second SDH assembly factor. Knockout of At5g67490 (sdhaf4) did not cause any phenotypic variation in seedlings but resulted in a decrease in both SDH activity and the succinate-dependent respiration rate as well as increased accumulation of succinate. Mass spectrometry analyses revealed stable levels of FAD-SDH1 in sdhaf4, together with increased levels of the FAD-SDH1 assembly factor, SDHAF2, and reduced levels of SDH2 compared with the wild type. Loss of SDHAF4 in sdhaf4 inhibited the formation of the SDH1/SDH2 intermediate, leading to the accumulation of soluble SDH1 in the mitochondrial matrix and reduced levels of SDH1 in the membrane. The increased levels of SDHAF2 suggest that the stabilization of soluble FAD-SDH1 depends on SDHAF2 availability. We conclude that SDHAF4 acts on FAD-SDH1 and promotes its assembly with SDH2, thereby stabilizing SDH2 and enabling its full assembly with SDH3/SDH4 to form the SDH complex.
Original languageEnglish
Pages (from-to)1439-1452
JournalPlant Physiology
Volume177
Issue number4
DOIs
Publication statusPublished - Aug 2018

Fingerprint

succinate dehydrogenase (quinone)
Flavin-Adenine Dinucleotide
Succinate Dehydrogenase
succinic acid
phenotypic variation
Succinic Acid
Arabidopsis
Arabidopsis thaliana
mass spectrometry
metabolism
seedlings
enzymes
Respiratory Rate
Seedlings
Mass Spectrometry
Membranes
Enzymes

Cite this

@article{2529601287e443a6a7f6ce0db2c9fa8f,
title = "An assembly factor promotes assembly of flavinated SDH1 into the succinate dehydrogenase complex",
abstract = "Succinate dehydrogenase (Complex II; SDH) plays an important role in mitochondrial respiratory metabolism. The SDH complex consists of four core subunits and multiple cofactors, which must be assembled correctly to ensure enzyme function. To date, only an assembly factor (SDHAF2) required for FAD insertion into subunit SDH1 has been identified in plants. Here, we report the identification of Arabidopsis (Arabidopsis thaliana) At5g67490 as a second SDH assembly factor. Knockout of At5g67490 (sdhaf4) did not cause any phenotypic variation in seedlings but resulted in a decrease in both SDH activity and the succinate-dependent respiration rate as well as increased accumulation of succinate. Mass spectrometry analyses revealed stable levels of FAD-SDH1 in sdhaf4, together with increased levels of the FAD-SDH1 assembly factor, SDHAF2, and reduced levels of SDH2 compared with the wild type. Loss of SDHAF4 in sdhaf4 inhibited the formation of the SDH1/SDH2 intermediate, leading to the accumulation of soluble SDH1 in the mitochondrial matrix and reduced levels of SDH1 in the membrane. The increased levels of SDHAF2 suggest that the stabilization of soluble FAD-SDH1 depends on SDHAF2 availability. We conclude that SDHAF4 acts on FAD-SDH1 and promotes its assembly with SDH2, thereby stabilizing SDH2 and enabling its full assembly with SDH3/SDH4 to form the SDH complex.",
author = "Katharina Belt and {Van Aken}, Olivier and Monika Murcha and Andrew Millar and Shaobai Huang",
year = "2018",
month = "8",
doi = "10.1104/pp.18.00320",
language = "English",
volume = "177",
pages = "1439--1452",
journal = "Plant Physiology (Online)",
issn = "0032-0889",
publisher = "American Society of Plant Biologists",
number = "4",

}

An assembly factor promotes assembly of flavinated SDH1 into the succinate dehydrogenase complex. / Belt, Katharina; Van Aken, Olivier; Murcha, Monika; Millar, Andrew; Huang, Shaobai.

In: Plant Physiology, Vol. 177, No. 4, 08.2018, p. 1439-1452.

Research output: Contribution to journalArticle

TY - JOUR

T1 - An assembly factor promotes assembly of flavinated SDH1 into the succinate dehydrogenase complex

AU - Belt, Katharina

AU - Van Aken, Olivier

AU - Murcha, Monika

AU - Millar, Andrew

AU - Huang, Shaobai

PY - 2018/8

Y1 - 2018/8

N2 - Succinate dehydrogenase (Complex II; SDH) plays an important role in mitochondrial respiratory metabolism. The SDH complex consists of four core subunits and multiple cofactors, which must be assembled correctly to ensure enzyme function. To date, only an assembly factor (SDHAF2) required for FAD insertion into subunit SDH1 has been identified in plants. Here, we report the identification of Arabidopsis (Arabidopsis thaliana) At5g67490 as a second SDH assembly factor. Knockout of At5g67490 (sdhaf4) did not cause any phenotypic variation in seedlings but resulted in a decrease in both SDH activity and the succinate-dependent respiration rate as well as increased accumulation of succinate. Mass spectrometry analyses revealed stable levels of FAD-SDH1 in sdhaf4, together with increased levels of the FAD-SDH1 assembly factor, SDHAF2, and reduced levels of SDH2 compared with the wild type. Loss of SDHAF4 in sdhaf4 inhibited the formation of the SDH1/SDH2 intermediate, leading to the accumulation of soluble SDH1 in the mitochondrial matrix and reduced levels of SDH1 in the membrane. The increased levels of SDHAF2 suggest that the stabilization of soluble FAD-SDH1 depends on SDHAF2 availability. We conclude that SDHAF4 acts on FAD-SDH1 and promotes its assembly with SDH2, thereby stabilizing SDH2 and enabling its full assembly with SDH3/SDH4 to form the SDH complex.

AB - Succinate dehydrogenase (Complex II; SDH) plays an important role in mitochondrial respiratory metabolism. The SDH complex consists of four core subunits and multiple cofactors, which must be assembled correctly to ensure enzyme function. To date, only an assembly factor (SDHAF2) required for FAD insertion into subunit SDH1 has been identified in plants. Here, we report the identification of Arabidopsis (Arabidopsis thaliana) At5g67490 as a second SDH assembly factor. Knockout of At5g67490 (sdhaf4) did not cause any phenotypic variation in seedlings but resulted in a decrease in both SDH activity and the succinate-dependent respiration rate as well as increased accumulation of succinate. Mass spectrometry analyses revealed stable levels of FAD-SDH1 in sdhaf4, together with increased levels of the FAD-SDH1 assembly factor, SDHAF2, and reduced levels of SDH2 compared with the wild type. Loss of SDHAF4 in sdhaf4 inhibited the formation of the SDH1/SDH2 intermediate, leading to the accumulation of soluble SDH1 in the mitochondrial matrix and reduced levels of SDH1 in the membrane. The increased levels of SDHAF2 suggest that the stabilization of soluble FAD-SDH1 depends on SDHAF2 availability. We conclude that SDHAF4 acts on FAD-SDH1 and promotes its assembly with SDH2, thereby stabilizing SDH2 and enabling its full assembly with SDH3/SDH4 to form the SDH complex.

U2 - 10.1104/pp.18.00320

DO - 10.1104/pp.18.00320

M3 - Article

VL - 177

SP - 1439

EP - 1452

JO - Plant Physiology (Online)

JF - Plant Physiology (Online)

SN - 0032-0889

IS - 4

ER -