Biosensor reveals multiple sources for mitochondrial NAD+

Xiaolu A. Cambronne; Melissa L. Stewart; Dongho Kim; Amber M. Jones-Brunette; Rory K. Morgan; David L. Farrens; Michael S. Cohen; Richard H. Goodman

doi:10.1126/science.aad5168

Biosensor reveals multiple sources for mitochondrial NAD+

Xiaolu A. Cambronne, Melissa L. Stewart, Dongho Kim, Amber M. Jones-Brunette, Rory K. Morgan, David L. Farrens, Michael S. Cohen, Richard H. Goodman

Research output: Contribution to journal › Article › peer-review

278 Scopus citations

Abstract

Nicotinamide adenine dinucleotide (NAD⁺) is an essential substrate for sirtuins and poly(adenosine diphosphate-ribose) polymerases (PARPs), which are NAD⁺-consuming enzymes localized in the nucleus, cytosol, and mitochondria. Fluctuations in NAD⁺ concentrations within these subcellular compartments are thought to regulate the activity of NAD⁺-consuming enzymes; however, the challenge in measuring compartmentalized NAD⁺ in cells has precluded direct evidence for this type of regulation.We describe the development of a genetically encoded fluorescent biosensor for directly monitoring free NAD⁺ concentrations in subcellular compartments. We found that the concentrations of free NAD⁺ in the nucleus, cytoplasm, and mitochondria approximate the Michaelis constants for sirtuins and PARPs in their respective compartments. Systematic depletion of enzymes that catalyze the final step of NAD⁺ biosynthesis revealed cell-specific mechanisms for maintaining mitochondrial NAD⁺ concentrations.

Original language	English (US)
Pages (from-to)	1474-1477
Number of pages	4
Journal	Science
Volume	352
Issue number	6292
DOIs	https://doi.org/10.1126/science.aad5168
State	Published - Jun 17 2016

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title = "Biosensor reveals multiple sources for mitochondrial NAD+",

abstract = "Nicotinamide adenine dinucleotide (NAD+) is an essential substrate for sirtuins and poly(adenosine diphosphate-ribose) polymerases (PARPs), which are NAD+-consuming enzymes localized in the nucleus, cytosol, and mitochondria. Fluctuations in NAD+ concentrations within these subcellular compartments are thought to regulate the activity of NAD+-consuming enzymes; however, the challenge in measuring compartmentalized NAD+ in cells has precluded direct evidence for this type of regulation.We describe the development of a genetically encoded fluorescent biosensor for directly monitoring free NAD+ concentrations in subcellular compartments. We found that the concentrations of free NAD+ in the nucleus, cytoplasm, and mitochondria approximate the Michaelis constants for sirtuins and PARPs in their respective compartments. Systematic depletion of enzymes that catalyze the final step of NAD+ biosynthesis revealed cell-specific mechanisms for maintaining mitochondrial NAD+ concentrations.",

author = "Cambronne, {Xiaolu A.} and Stewart, {Melissa L.} and Dongho Kim and Jones-Brunette, {Amber M.} and Morgan, {Rory K.} and Farrens, {David L.} and Cohen, {Michael S.} and Goodman, {Richard H.}",

note = "Funding Information: This work was supported by the Pew Charitable Trust and the NIH (grants NS088629, UL1TR000128, P30NS061800, and P30CA069533).",

year = "2016",

month = jun,

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doi = "10.1126/science.aad5168",

language = "English (US)",

volume = "352",

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journal = "Science",

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T1 - Biosensor reveals multiple sources for mitochondrial NAD+

AU - Cambronne, Xiaolu A.

AU - Stewart, Melissa L.

AU - Kim, Dongho

AU - Jones-Brunette, Amber M.

AU - Morgan, Rory K.

AU - Farrens, David L.

AU - Cohen, Michael S.

AU - Goodman, Richard H.

N1 - Funding Information: This work was supported by the Pew Charitable Trust and the NIH (grants NS088629, UL1TR000128, P30NS061800, and P30CA069533).

PY - 2016/6/17

Y1 - 2016/6/17

N2 - Nicotinamide adenine dinucleotide (NAD+) is an essential substrate for sirtuins and poly(adenosine diphosphate-ribose) polymerases (PARPs), which are NAD+-consuming enzymes localized in the nucleus, cytosol, and mitochondria. Fluctuations in NAD+ concentrations within these subcellular compartments are thought to regulate the activity of NAD+-consuming enzymes; however, the challenge in measuring compartmentalized NAD+ in cells has precluded direct evidence for this type of regulation.We describe the development of a genetically encoded fluorescent biosensor for directly monitoring free NAD+ concentrations in subcellular compartments. We found that the concentrations of free NAD+ in the nucleus, cytoplasm, and mitochondria approximate the Michaelis constants for sirtuins and PARPs in their respective compartments. Systematic depletion of enzymes that catalyze the final step of NAD+ biosynthesis revealed cell-specific mechanisms for maintaining mitochondrial NAD+ concentrations.

AB - Nicotinamide adenine dinucleotide (NAD+) is an essential substrate for sirtuins and poly(adenosine diphosphate-ribose) polymerases (PARPs), which are NAD+-consuming enzymes localized in the nucleus, cytosol, and mitochondria. Fluctuations in NAD+ concentrations within these subcellular compartments are thought to regulate the activity of NAD+-consuming enzymes; however, the challenge in measuring compartmentalized NAD+ in cells has precluded direct evidence for this type of regulation.We describe the development of a genetically encoded fluorescent biosensor for directly monitoring free NAD+ concentrations in subcellular compartments. We found that the concentrations of free NAD+ in the nucleus, cytoplasm, and mitochondria approximate the Michaelis constants for sirtuins and PARPs in their respective compartments. Systematic depletion of enzymes that catalyze the final step of NAD+ biosynthesis revealed cell-specific mechanisms for maintaining mitochondrial NAD+ concentrations.

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JO - Science

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Biosensor reveals multiple sources for mitochondrial NAD+

Abstract

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