Synthesizing and Salvaging NAD: Lessons Learned from
The essential coenzyme nicotinamide adenine dinucleotide (NAD+) plays important roles in metabolic reactions and cell regulation in all organisms. Bacteria, fungi, plants, and animals use different pathways to synthesize NAD+. Our molecular and genetic data demonstrate that in the unicellular green alga Chlamydomonas NAD+ is synthesized from aspartate (de novo synthesis), as in plants, or nicotinamide, as in mammals (salvage synthesis). The de novo pathway requires five different enzymes: L-aspartate oxidase (ASO), quinolinate synthetase (QS), quinolate phosphoribosyltransferase (QPT), nicotinate/nicotinamide mononucleotide adenylyltransferase (NMNAT), and NAD+ synthetase (NS). Sequence similarity searches, gene isolation and sequencing of mutant loci indicate that mutations in each enzyme result in a nicotinamide-requiring mutant phenotype in the previously isolated nic mutants. We rescued the mutant phenotype by the introduction of BAC DNA (nic2-1 and nic13-1) or plasmids with cloned genes (nic1-1 and nic15-1) into the mutants. NMNAT, which is also in the de novo pathway, and nicotinamide phosphoribosyltransferase (NAMPT) constitute the nicotinamide-dependent salvage pathway. A mutation in NAMPT (npt1-1) has no obvious growth defect and is not nicotinamide-dependent. However, double mutant strains with the npt1-1 mutation and any of the nic mutations are inviable. When the de novo pathway is inactive, the salvage pathway is essential to Chlamydomonas for the synthesis of NAD+. A homolog of the human SIRT6-like gene, SRT2, is upregulated in the NS mutant, which shows a longer vegetative life span than wild-type cells. Our results suggest that Chlamydomonas is an excellent model system to study NAD+ metabolism and cell longevity.
Vyšlo v časopise:
Synthesizing and Salvaging NAD: Lessons Learned from. PLoS Genet 6(9): e32767. doi:10.1371/journal.pgen.1001105
Kategorie:
Research Article
prolekare.web.journal.doi_sk:
https://doi.org/10.1371/journal.pgen.1001105
Souhrn
The essential coenzyme nicotinamide adenine dinucleotide (NAD+) plays important roles in metabolic reactions and cell regulation in all organisms. Bacteria, fungi, plants, and animals use different pathways to synthesize NAD+. Our molecular and genetic data demonstrate that in the unicellular green alga Chlamydomonas NAD+ is synthesized from aspartate (de novo synthesis), as in plants, or nicotinamide, as in mammals (salvage synthesis). The de novo pathway requires five different enzymes: L-aspartate oxidase (ASO), quinolinate synthetase (QS), quinolate phosphoribosyltransferase (QPT), nicotinate/nicotinamide mononucleotide adenylyltransferase (NMNAT), and NAD+ synthetase (NS). Sequence similarity searches, gene isolation and sequencing of mutant loci indicate that mutations in each enzyme result in a nicotinamide-requiring mutant phenotype in the previously isolated nic mutants. We rescued the mutant phenotype by the introduction of BAC DNA (nic2-1 and nic13-1) or plasmids with cloned genes (nic1-1 and nic15-1) into the mutants. NMNAT, which is also in the de novo pathway, and nicotinamide phosphoribosyltransferase (NAMPT) constitute the nicotinamide-dependent salvage pathway. A mutation in NAMPT (npt1-1) has no obvious growth defect and is not nicotinamide-dependent. However, double mutant strains with the npt1-1 mutation and any of the nic mutations are inviable. When the de novo pathway is inactive, the salvage pathway is essential to Chlamydomonas for the synthesis of NAD+. A homolog of the human SIRT6-like gene, SRT2, is upregulated in the NS mutant, which shows a longer vegetative life span than wild-type cells. Our results suggest that Chlamydomonas is an excellent model system to study NAD+ metabolism and cell longevity.
Zdroje
1. BakkerBM
OverkampKM
MarisAJA
KotterP
LuttikMAH
2001 Stoichiometry and compartmentation of NADH metabolism in Saccharomyces cerevisiae. FEMS Microbiol Rev 25 15 37
2. BelenkyP
BoganKL
BrennerC
2007 NAD+ metabolism in health and disease. Trends Biochem Sci 32 12 19
3. VrablikTL
HuangL
LangeSE
Hanna-RoseW
2009 Nicotinamidase modulation of NAD+ biosynthesis and nicotinamide levels separately affect reproductive development and cell survival in C. elegans. Development 136 3637 3746
4. TannyJC
DowdGJ
HuangJ
HilzH
MoazedD
1999 An enzymatic activity in the yeast Sir2 protein that is essential for gene silencing. Cell 99 735 745
5. OberdoerfferP
MichanS
McVayM
MostoslavskyR
VannJ
2008 SIRT1 redistribution on chromatin promotes genomic stability but alters gene expression during aging. Cell 135 907 918
6. InoueT
HiratsukaM
OsakiM
OshimuraM
2007 The molecular biology of mammalian SIRT proteins: SIRT2 in cell cycle regulation. Cell Cycle 6 1011 1018
7. MostoslavskyR
ChuaKF
LombardDB
PangWW
FischerMR
2006 Genomic instability and aging-like phenotype in the absence of mammalian SIRT6. Cell 124 315 329
8. ZhongL
D'UrsoA
ToiberD
SebastianC
HenryRE
2010 The histone deacetylase Sirt6 regulates glucose homeostasis via Hif1 [alpha]. Cell 140 280 293
9. LuoJ
NikolaevAY
ImaiS
ChenD
SuF
2001 Negative control of p53 by Sir2a promotes cell survival under stress. Cell 107 137 148
10. FryeRA
2000 Phylogenetic classification of prokaryotic and eukaryotic Sir2-like proteins. Biochem Biophys Res Commun 273 793 798
11. GreissS
GartnerA
2009 Sirtuin/Sir2 phylogeny, evolutionary considerations and structural conservation. Mol Cells 28 407 415
12. MichishitaE
McCordRA
BerberE
KioiM
Padilla-NashH
2008 SIRT6 is a histone H3 lysine 9 deacetylase that modulates telomeric chromatin. Nature 452 492 496
13. FordE
VoitR
LisztG
MaginC
GrummtI
2006 Mammalian Sir2 homolog SIRT7 is an activator of RNA polymerase I transcription. Genes Dev 20 1075 1080
14. GrobA
RousselP
WrightJ
McStayB
Hernandez-VerdunD
2009 Involvement of SIRT7 in resumption of rDNA transcription at the exit from mitosis. J Cell Sci 122 489
15. MerchantSS
ProchnikSE
VallonO
HarrisEH
KarpowiczSJ
2007 The Chlamydomonas genome reveals the evolution of key animal and plant functions. Science 318 245 250
16. ArchibaldJM
2009 GENOMICS: Green Evolution, Green Revolution. Science 324 191 192
17. KatohA
HashimotoT
2004 Molecular biology of pyridine nucleotide and nicotine biosynthesis. Front Biosci 9 1577 1586
18. EversoleRA
1956 Biochemical mutants of Chlamydomonas reinhardi. Am J Botany 43 404 407
19. HarrisEH
1989 The Chlamydomonas Sourcebook: a comprehensive guide to biology and laboratory use San Diego Academic Press xiv, 780
20. FerrisPJ
1995 Localization of the nic-7, ac-29 and thi-10 genes within the mating-type locus of Chlamydomonas reinhardtii. Genetics 141 543 549
21. FerrisPJ
ArmbrustEV
GoodenoughUW
2002 Genetic structure of the mating-type locus of Chlamydomonas reinhardtii. Genetics 160 181 200
22. DutcherSK
PowerJ
GallowayRE
PorterME
1991 Reappraisal of the genetic map of Chlamydomonas reinhardtii. J Hered 82 295 301
23. EbersoldWT
LevineRP
1959 A genetic analysis of linkage group I of Chlamydomonas reinhardi. Z Vererbungsl 90 74 82
24. EbersoldWT
LevineRP
LevineEE
OlmstedMA
1962 Linkage maps in Chlamydomonas reinhardi. Genetics 47 531 543
25. SmythRD
MartinekGW
EbersoldWT
1975 Linkage of six genes in Chlamydomonas reinhardtii and the construction of linkage test strains. J Bacteriol 124 1615 1617
26. WoolleyD
1945 Production of nicotinic acid deficiency with 3-acetylpyridine, the ketone analogue of nicotinic acid. J Biol Chem 157 455
27. WoolleyD
1946 Reversal by tryptophane of the biological effects of 3-acetylpyridine. J Biol Chem 162 179
28. KatohA
UenoharaK
AkitaM
HashimotoT
2006 Early steps in the biosynthesis of NAD in Arabidopsis start with aspartate and occur in the plastid. Plant Physiol 141 851 857
29. SchippersJHM
Nunes-NesiA
ApetreiR
HilleJ
FernieAR
2008 The Arabidopsis onset of leaf death5 mutation of quinolinate synthase affects nicotinamide adenine dinucleotide biosynthesis and causes early ageing. Plant Cell 20 2909 2925
30. PazourGJ
KoutoulisA
BenashskiSE
DickertBL
ShengH
1999 LC2, the Chlamydomonas homologue of the t complex-encoded protein Tctex2, is essential for outer dynein arm assembly. Mol Biol Cell 10 3507 3520
31. KathirP
LaVoieM
BrazeltonWJ
HaasNA
LefebvrePA
2003 Molecular map of the Chlamydomonas reinhardtii nuclear genome. Eukaryot Cell 2 362 379
32. LiuH
WoznicaK
CattonG
CrawfordA
BottingN
2007 Structural and kinetic characterization of quinolinate phosphoribosyltransferase (hQPRTase) from Homo sapiens. J Mol Biol 373 755 763
33. KwanAL
LiL
KulpDC
DutcherSK
StormoGD
2009 Improving gene-finding in Chlamydomonas reinhardtii: GreenGenie 2. BMC Genomics 10 e210
34. PerroneCA
MysterSH
BowerR
O'TooleET
PorterME
2000 Insights into the structural organization of the I1 inner arm dynein from a domain analysis of the 1-b dynein heavy chain. Mol Biol Cell 11 2297 2313
35. SizovaI
FuhrmannM
HegemannP
2001 A Streptomyces rimosus aphVIII gene coding for a new type phosphotransferase provides stable antibiotic resistance to Chlamydomonas reinhardtii. Gene 277 221 229
36. HarrisE
2009 The Chlamydomonas Sourcebook; Second Edition Academic Press
37. RaffaelliN
LorenziT
MarianiPL
EmanuelliM
AmiciA
1999 The Escherichia coli NadR regulator is endowed with nicotinamide mononucleotide adenylyltransferase activity. J Bacteriol 181 5509 5511
38. GroseJH
BergthorssonU
RothJR
2005 Regulation of NAD synthesis by the trifunctional NadR protein of Salmonella enterica. J Bacteriol 187 2774 2782
39. NakahataY
SaharS
AstaritaG
KaluzovaM
Sassone-CorsiP
2009 Circadian control of the NAD+ salvage pathway by CLOCK-SIRT1. Science 324 654 657
40. TissenbaumH
GuarenteL
2001 Increased dosage of a sir-2 gene extends lifespan in Caenorhabditis elegans. Nature 410 227 230
41. LinS
DefossezP
GuarenteL
2000 Requirement of NAD and SIR2 for life-span extension by calorie restriction in Saccharomyces cerevisiae. Science 289 2126
42. HuangL
SunQ
QinF
LiC
ZhaoY
2007 Down-regulation of a SILENT INFORMATION REGULATOR2-related histone deacetylase gene, OsSRT1, induces DNA fragmentation and cell death in rice. Plant Physiol 144 1508
43. DutcherSK
TrabucoEC
1998 The UNI3 gene is required for assembly of basal bodies of Chlamydomonas and encodes delta-tubulin, a new member of the tubulin superfamily. Mol Biol Cell 9 1293 1308
44. ZhangQ
WangS-Y
FleurielC
LeprinceD
RocheleauJV
2007 Metabolic regulation of SIRT1 transcription via a HIC1:CtBP corepressor complex. Proc Natl Acad Sci U S A 104 829 833
45. FragaMF
EstellerM
2007 Epigenetics and aging: the targets and the marks. Trends Genet 23 413 418
46. YangH
YangT
BaurJA
PerezE
MatsuiT
2007 Nutrient-sensitive mitochondrial NAD+ levels dictate cell survival. Cell 130 1095 1107
47. ZhaiRG
ZhangF
HiesingerPR
CaoY
HaueterCM
2008 NAD synthase NMNAT acts as a chaperone to protect against neurodegeneration. Nature 452 887 891
48. RutterJ
ReickM
WuLC
McKnightSL
2001 Regulation of clock and NPAS2 DNA binding by the redox state of NAD cofactors. Science 293 510 514
49. VanderauweraS
De BlockM
Van de SteeneN
van de CotteB
MetzlaffM
2007 Silencing of poly(ADP-ribose) polymerase in plants alters abiotic stress signal transduction. Proc Natl Acad Sci U S A 104 15150 15155
50. van der VeerE
HoC
O'NeilC
BarbosaN
ScottR
2007 Extension of human cell lifespan by nicotinamide phosphoribosyltransferase. J Biol Chem 282 10841 10845
51. VaqueroA
SternglanzR
ReinbergD
2007 NAD+-dependent deacetylation of H4 lysine 16 by class III HDACs. Oncogene 26 5505 5520
52. KohanskiMA
DwyerDJ
HayeteB
LawrenceCA
CollinsJJ
2007 A common mechanism of cellular death induced by bactericidal antibiotics. Cell 130 797 810
53. RongvauxA
AndrisF
Van GoolF
LeoO
2003 Reconstructing eukaryotic NAD metabolism. Bioessays 25 683 690
54. KurnasovO
GoralV
ColabroyK
GerdesS
AnanthaS
2003 NAD biosynthesis identification of the tryptophan to quinolinate pathway in bacteria. Chem Biol 10 1195 1204
55. KobayashiK
EhrlichS
AlbertiniA
AmatiG
AndersenK
2003 Essential Bacillus subtilis genes. Proc Natl Acad Sci U S A 100 4678 4683
56. ImaiS
2009 Nicotinamide phosphoribosyltransferase (Nampt): a link between NAD biology, metabolism, and diseases. Current Pharmaceutical Design 15 20 28
57. HashidaSN
TakahashiH
Kawai-YamadaM
UchimiyaH
2007 Arabidopsis thaliana nicotinate/nicotinamide mononucleotide adenyltransferase (AtNMNAT) is required for pollen tube growth. Plant J 49 694 703
58. RevolloJR
KörnerA
MillsKF
SatohA
WangT
2007 Nampt/PBEF/Visfatin regulates insulin secretion in [beta] cells as a systemic NAD biosynthetic enzyme. Cell Metab 6 363 375
59. ShinDH
OganesyaN
JancarikJ
YokotaH
KimR
KimSH
2005 Crystal structure of a nicotinate phosphoribosyltransferase from Thermoplasma acidophilum. J Biol Chem 280 18326 18335
60. ChappieJS
CànavesJM
HanGW
RifeCL
XuQ
2005 The Structure of a Eukaryotic Nicotinic Acid Phosphoribosyltransferase Reveals Structural Heterogeneity among Type II PRTases. Structure 13 1385 1396
61. KirkDL
1998 Volvox: A Search for the Molecular and Genetic Origins of Multicellularity and Cellular Differentiation Cambridge University Press 399
62. PalenikB
GrimwoodJ
AertsA
RouzeP
SalamovA
2007 The tiny eukaryote Ostreococcus provides genomic insights into the paradox of plankton speciation. Proc Natl Acad Sci U S A 104 7705 7710
63. DerelleE
FerrazC
RombautsS
RouzeP
WordenAZ
2006 Genome analysis of the smallest free-living eukaryote Ostreococcus tauri unveils many unique features. Proc Natl Acad Sci U S A 103 11647 11652
64. PeersG
NiyogiKK
2008 Pond scum genomics: the genomes of Chlamydomonas and Ostreococcus. Plant Cell 20 502 507
65. KingN
WestbrookMJ
YoungSL
KuoA
AbedinM
2008 The genome of the choanoflagellate Monosiga brevicollis and the origin of metazoans. Nature 451 783 788
66. SmallI
PeetersN
LegeaiF
LurinC
2004 Predotar: A tool for rapidly screening proteomes for N-terminal targeting sequences. Proteomics 4 1581 1590
67. EmanuelssonO
NielsenH
BrunakS
von HeijneG
2000 Predicting subcellular localization of proteins based on their N-terminal amino acid sequence. J Mol Biol 300 1005 1016
68. GaravagliaS
D'AngeloI
EmanuelliM
CarnevaliF
PierellaF
2002 Structure of human NMN adenylytransferase. A key nuclear enzyme for NAD homeostasis. J. Biol Chem 8 8524 8530
69. ZhaiRG
CaoY
HiesingerPR
ZhouY
MehtaSQ
2006 Drosophila NMNAT maintains neural integrity independent of its NAD synthesis activity. PLoS Biol 4 e416
70. NakamuraK
GowansCS
1965 Genetic control of nicotinic acid metabolism in Chlamydomonas eugametos. Genetics 51 931 935
71. BieganowskiP
BrennerC
2004 Discoveries of nicotinamide riboside as a nutrient and conserved NRK genes establish a Preiss-Handler independent route to NAD+ in fungi and humans. Cell 117 495 502
72. BelenkyP
RacetteFG
BoganKL
McClureJM
SmithJS
2007 Nicotinamide riboside promotes Sir2 silencing and extends lifespan via Nrk and Urh1/Pnp1/Meu1 pathways to NAD+. Cell 129 473 484
73. BelenkyP
ChristensenKC
GazzanigaF
PletnevAA
BrennerC
2009 Nicotinamide riboside and nicotinic acid riboside salvage in fungi and mammals: Quantitative basis for Urh1 and purine nucleoside phosphorylase function in NAD+ metabolism. J Biol Chem 284 158 164
74. KaeberleinM
McVeyM
GuarenteL
1999 The SIR2/3/4 complex and SIR2 alone promote longevity in Saccharomyces cerevisiae by two different mechanisms. Genes Dev 13 2570
75. RoginaB
HelfandSL
2004 Sir2 mediates longevity in the fly through a pathway related to calorie restriction. Proc Natl Acad Sci USA 101 15998 16003
76. CohenH
MillerC
BittermanK
WallN
HekkingB
2004 Calorie restriction promotes mammalian cell survival by inducing the SIRT1 deacetylase. Science 305 390
77. SchulzTJ
ZarseK
VoigtA
UrbanN
BirringerM
2007 Glucose restriction extends Caenorhabditis elegans life span by inducing mitochondrial respiration and increasing oxidative stress. Cell Metab 6 280 293
78. ChengJ
TürkelN
HematiN
FullerMT
HuntAJ
2008 Centrosome misorientation reduces stem cell division during ageing. Nature 456 599 604
79. WangX
TsaiJW
ImaiJH
LianWN
ValleeRB
2009 Asymmetric centrosome inheritance maintains neural progenitors in the neocortex. Nature 461 947 955
80. AsherG
GatfieldD
StratmannM
ReinkeH
DibnerC
2008 SIRT1 regulates circadian clock gene expression through PER2 deacetylation. Cell 134 317 328
81. MatsuoT
OkamotoK
OnaiK
NiwaY
ShimogawaraK
2008 A systematic forward genetic analysis identified components of the Chlamydomonas circadian system. Genes Dev 22 918 930
82. LuxFG
DutcherSK
1991 Genetic interactions at the FLA10 locus: suppressors and synthetic phenotypes that affect the cell cycle and flagellar function in Chlamydomonas reinhardtii. Genetics 128 549 561
83. LarkinMA
BlackshieldsG
BrownNP
ChennaR
McGettiganPA
2007 Clustal W and Clustal X version 2.0. Bioinformatics 23 2947 2948
84. LinH
GoodenoughUW
2007 Gametogenesis in the Chlamydomonas reinhardtii minus mating type is controlled by two genes, MID and MTD1. Genetics 176 913 925
85. BarnesWM
1994 PCR amplification of up to 35 kb DNA with high fidelity and high yield from lambda bacteriophage templates. Proc Natl Acad Sci 91 2216 2220
86. NewmanSM
BoyntonJE
GillhamNW
Randolph-AndersonBL
JohnsonAM
1990 Transformation of chloroplast ribosomal RNA genes in Chlamydomonas: molecular and genetic characterization of integration events. Genetics 126 875 888
87. IominiC
LiL
EsparzaJM
DutcherSK
2009 Retrograde IFT mutants identify complex A proteins with multiple genetic interactions in Chlamydomonas reinhardtii. Genetics 183 885 896
88. FangSC
De Los ReyesC
UmenJG
2006 Cell size checkpoint control by the retinoblastoma tumor suppressor pathway. PLoS Genet 2 e167
89. EngelkeDR
KrikosA
BruckME
GinsburgD
1990 Purification of Thermus aquaticus DNA polymerase expressed in Escherichia coli. Analytical Biochemistry 191 396 400
90. NelsonJA
SavereidePB
LefebvrePA
1994 The CRY1 gene in Chlamydomonas reinhardtii: structure and use as a dominant selectable marker for nuclear transformation. Mol Cell Biol 14 4011 4019
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Genetika Reprodukčná medicínaČlánok vyšiel v časopise
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