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PMID:27694803

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Citation

Lin, Z, Dong, M, Zhang, Y, Lee, EA and Lin, H' (2016) Cbr1 is a Dph3 reductase required for the tRNA wobble uridine modification. Nat. Chem. Biol. '

Abstract

Diphthamide and the tRNA wobble uridine modifications both require diphthamide biosynthesis 3 (Dph3) protein as an electron donor for the iron-sulfur clusters in their biosynthetic enzymes. Here, using a proteomic approach, we identified Saccharomyces cerevisiae cytochrome b5 reductase (Cbr1) as a NADH-dependent reductase for Dph3. The NADH- and Cbr1-dependent reduction of Dph3 may provide a regulatory linkage between cellular metabolic state and protein translation.

Links

PubMed Online version:10.1038/nchembio.2190

Keywords


Significance

Annotations

Gene product Qualifier GO Term Evidence Code with/from Aspect Extension Notes Status

YEAST:NCB5R

involved_in

GO:0017183: peptidyl-diphthamide biosynthetic process from peptidyl-histidine

ECO:0000315: mutant phenotype evidence used in manual assertion

P

Seeded From UniProt

complete

YEAST:NCB5R

involved_in

GO:0002926: tRNA wobble base 5-methoxycarbonylmethyl-2-thiouridinylation

ECO:0000315: mutant phenotype evidence used in manual assertion

P

Seeded From UniProt

complete

YEAST:DPH3

involved_in

GO:0017183: peptidyl-diphthamide biosynthetic process from peptidyl-histidine

ECO:0000315: mutant phenotype evidence used in manual assertion

P

Seeded From UniProt

complete

YEAST:DPH3

involved_in

GO:0002926: tRNA wobble base 5-methoxycarbonylmethyl-2-thiouridinylation

ECO:0000315: mutant phenotype evidence used in manual assertion

P

Seeded From UniProt

complete

YEAST:NCPR

GO:0003959: NADPH dehydrogenase activity

ECO:0000314:

F

The author cloned and overexpressed Ncp1 and monitored the reduction of Dph3 by Dph3's absorbance at 488nm. The author found that Ncp1 reduced Dph3 after the addition of NADPH to the reaction tube (Supplementary figure 5C).

complete
CACAO 12191

YEAST:NCB5R

GO:0003954: NADH dehydrogenase activity

ECO:0000314:

F

The author expressed Cbr1 and monitored the reduction of Dph3 through Dph3's absorption at 488nm. The author then added NADH to the reaction tube and noticed that Cbr1 rapidly reduced Dph3 (this is shown is figure 1b). However, the author also found that Cbr1 did not reduce Dph3 upon addition of NADPH (supplemental figure 3).

complete
CACAO 12135

YEAST:MCR1

GO:0003954: NADH dehydrogenase activity

ECO:0000314:

F

The author cloned and overexpressed Mcr1 and monitored the reduction of Dph3 by Dph3's absorbance at 488nm. The author found that Mcr1 reduced Dph3 after the addition of NADH to the reaction tube (Supplementary figure 5A).

complete
CACAO 12136

Notes

See also

References

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