Quantification of protein copy number in yeast: the NAD+ metabolome

Szu-Chieh Mei; Charles Brenner

doi:10.1371/journal.pone.0106496

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Quantification of protein copy number in yeast: the NAD+ metabolome

Journal article

Open access

Peer reviewed

Quantification of protein copy number in yeast: the NAD+ metabolome

Szu-Chieh Mei and Charles Brenner

PloS one, Vol.9(9), e106496

2014

DOI: 10.1371/journal.pone.0106496

PMCID: PMC4154715

PMID: 25188219

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Abstract

Saccharomyces cerevisiae is calorie-restricted by lowering glucose from 2% to 0.5%. Under low glucose conditions, replicative lifespan is extended in a manner that depends on the NAD+-dependent protein lysine deacetylase Sir2 and NAD+ salvage enzymes. Because NAD+ is required for glucose utilization and Sir2 function, it was postulated that glucose levels alter the levels of NAD+ metabolites that tune Sir2 function. Though NAD+ precursor vitamins, which increase the levels of all NAD+ metabolites, can extend yeast replicative lifespan, glucose restriction does not significantly change the levels or ratios of intracellular NAD+ metabolites. To test whether glucose restriction affects protein copy numbers, we developed a technology that combines the measurement of Urh1 specific activity and quantification of relative expression between Urh1 and any other protein. The technology was applied to obtain the protein copy numbers of enzymes involved in NAD+ metabolism in rich and synthetic yeast media. Our data indicated that Sir2 and Pnc1, two enzymes that sequentially convert NAD+ to nicotinamide and then to nicotinic acid, are up-regulated by glucose restriction in rich media, and that Pnc1 alone is up-regulated in synthetic media while levels of all other enzymes are unchanged. These data suggest that production or export of nicotinic acid might be a connection between NAD+ and calorie restriction-mediated lifespan extension in yeast.

Saccharomyces cerevisiae - metabolism

Niacinamide - metabolism

Nicotinamidase - metabolism

Silent Information Regulator Proteins, Saccharomyces cerevisiae - metabolism

Saccharomyces cerevisiae Proteins - metabolism

Metabolome

NAD - metabolism

Niacin - metabolism

Sirtuin 2 - metabolism

Details

Title: Subtitle: Quantification of protein copy number in yeast: the NAD+ metabolome
Creators: Szu-Chieh Mei - Department of Biochemistry, Carver College of Medicine, University of Iowa, Iowa City, Iowa, United States of America
Charles Brenner - Department of Biochemistry, Carver College of Medicine, University of Iowa, Iowa City, Iowa, United States of America
Resource Type: Journal article
Publication Details: PloS one, Vol.9(9), e106496
DOI: 10.1371/journal.pone.0106496
PMID: 25188219
PMCID: PMC4154715
NLM abbreviation: PLoS One
ISSN: 1932-6203
eISSN: 1932-6203
Publisher: United States
Language: English
Date published: 2014
Academic Unit: Biochemistry and Molecular Biology; Internal Medicine
Record Identifier: 9983788597702771

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