Flavinylation and Assembly of Succinate Dehydrogenase Are Dependent on the C-terminal Tail of the Flavoprotein Subunit

Kim, Hyung J.; Jeong, Mi Young; Na, Un; Winge, Dennis R.

doi:10.1074/jbc.m112.405704

Cited by 39 publications

(77 citation statements)

References 29 publications

Supporting

Mentioning

Contrasting

Unclassified

Order By: Relevance

“…Eukaryotic Sdh5 binds to two Arg residues in the C-terminal tail of Sdh1. 16 This site is distal to the location of FAD in Sdh1, which would support a mechanism involving the induction of long-range conformational changes in Sdh1 induced by Sdh5.…”

Section: ■ Discussionmentioning

confidence: 98%

See 1 more Smart Citation

The Conserved RGxxE Motif of the Bacterial FAD Assembly Factor SdhE Is Required for Succinate Dehydrogenase Flavinylation and Activity

McNeil

Fineran

2013

Biochemistry

View full text Add to dashboard Cite

Succinate dehydrogenase (SDH) is an important respiratory enzyme that plays a critical role in the generation of energy in the majority of eukaryotes, bacteria, and archaea. The activity of SDH is dependent on the covalent attachment of the redox cofactor FAD to the flavoprotein subunit SdhA. In the Gram-negative bacteria Escherichia coli and Serratia sp. ATCC 39006, the covalent attachment of FAD to SdhA is dependent on the FAD assembly factor SdhE (YgfY). Although mechanisms have been proposed, experimental evidence that elucidates the molecular details of SdhE-mediated flavinylation are scarce. In this study, truncation and alanine swap mutagenesis of SdhE identified a highly conserved RGxxE motif that was important for SdhE function. Interestingly, RGxxE site-directed variants were not impaired in terms of protein folding or interactions with SdhA. Purification and analysis of SdhA from different mutant backgrounds demonstrated that SdhE interacts with and flavinylates folded SdhA without a requirement for the assembly of the entire SDH complex. SdhA was also partially active in the absence of SdhE, suggesting that SdhA is able to attach FAD through an inefficient autocatalytic mechanism. The results presented are of widespread relevance because SdhE and SDH are required for bacterial pathogenesis and mutations in the eukaryotic homologues of SdhE and SDH are associated with cancer in humans.

show abstract

Section: ■ Discussionmentioning

confidence: 98%

“…4 His-SdhA that was purified from ΔsdhCDAB::Kan contained covalently bound FAD ( Figure 1A). 16,26 In conclusion, SdhE flavinylates SdhA in the absence of other SDH complex proteins.…”

Section: ■ Experimental Proceduresmentioning

confidence: 98%

The Conserved RGxxE Motif of the Bacterial FAD Assembly Factor SdhE Is Required for Succinate Dehydrogenase Flavinylation and Activity

McNeil

Fineran

2013

Biochemistry

View full text Add to dashboard Cite

show abstract

“…We observed that SDHAF1 patient-derived cells had lower SDHA protein levels compared to control cells and drastically reduced levels of flavinylated SDHA. Studies in S. cerevisiae previously demonstrated that deletion of Sdh2 (SDHB ortholog in human) led to significant diminution of covalent flavinylation of Sdh1 (SDHA ortholog) (Kim et al, 2012; Robinson and Lemire, 1996), though the molecular mechanism by which Sdh2 promotes efficient flavinylation in vivo is unclear (Kim and Winge, 2013). Our new results show that impaired biogenesis of SDHB in mammalian cells also correlates with reduced flavinylation of SDHA.…”

Section: Discussionmentioning

confidence: 99%

Disease-Causing SDHAF1 Mutations Impair Transfer of Fe-S Clusters to SDHB

et al. 2016

View full text Add to dashboard Cite

SUMMARY SDHAF1 mutations cause a rare mitochondrial complex II (CII) deficiency, which manifests as infantile leukoencephalopathy with elevated levels of serum and white matter succinate and lactate. Here, we demonstrate that SDHAF1 contributes to iron-sulfur (Fe-S) cluster incorporation into the Fe-S subunit of CII, SDHB. SDHAF1 transiently binds to aromatic peptides of SDHB through an arginine-rich region in its C-terminus, and specifically engages a Fe-S donor complex, consisting of the scaffold, holo-ISCU, and the co-chaperone-chaperone pair, HSC20-HSPA9, through an LYR motif near its N-terminal domain. Pathogenic mutations of SDHAF1 abrogate binding to SDHB, which impairs biogenesis of holo-SDHB and results in LONP1-mediated degradation of SDHB. Riboflavin treatment was found to ameliorate the neurologic condition of patients. We demonstrate that riboflavin enhances flavinylation of SDHA, and reduces levels of succinate and Hypoxia-Inducible Factor (HIF) -1α and -2α, explaining the favorable response of patients to riboflavin.

show abstract

“…To further analyze the conservation are essential for SDH1 flavinylation and influence the binding and stability of SDHAF2. 8 Kim et al 8 proposed that the Arg residues could have roles: (1) directly in SDHAF2 association, (2) in the recruitment and/or guidance of FAD and/or (3) in allowing succinate to the substrate site to catalyze the covalent flavinylation reaction. The SDH1 sequence is quite conserved across different species at the protein sequence level but still many differences are evident across kingdoms.…”

Section: Sdhaf2 Is Divergent At the Sequence Level But A Short Consermentioning

confidence: 99%

“…7 Recent evidence in yeast indicates that the C-terminal tail of SDH1 is important for both flavinylation of SDH1 and for this SDH1:SDHAF2 interaction. 8 In yeast, two C-terminal Arg residues (R582 and R638; indicated by red arrows in Fig. 2A) which are distant from the FAD binding site …”

Section: The C-terminal Tail Of Sdh1 Required For Sdhaf2mentioning

confidence: 99%

Sequence diversity and conservation in factors influencing succinate dehydrogenase flavinylation

Huang

Millar

2013

Plant Signaling & Behavior

View full text Add to dashboard Cite

Flavinylation and Assembly of Succinate Dehydrogenase Are Dependent on the C-terminal Tail of the Flavoprotein Subunit

Cited by 39 publications

References 29 publications

The Conserved RGxxE Motif of the Bacterial FAD Assembly Factor SdhE Is Required for Succinate Dehydrogenase Flavinylation and Activity

The Conserved RGxxE Motif of the Bacterial FAD Assembly Factor SdhE Is Required for Succinate Dehydrogenase Flavinylation and Activity

Disease-Causing SDHAF1 Mutations Impair Transfer of Fe-S Clusters to SDHB

Sequence diversity and conservation in factors influencing succinate dehydrogenase flavinylation

Contact Info

Product

Resources

About