Tracing compartmentalized NADPH metabolism in the cytosol and mitochondria of mammalian cells.

Abstract:

:Eukaryotic cells compartmentalize biochemical processes in different organelles, often relying on metabolic cycles to shuttle reducing equivalents across intracellular membranes. NADPH serves as the electron carrier for the maintenance of redox homeostasis and reductive biosynthesis, with separate cytosolic and mitochondrial pools providing reducing power in each respective location. This cellular organization is critical for numerous functions but complicates analysis of metabolic pathways using available methods. Here we develop an approach to resolve NADP(H)-dependent pathways present within both the cytosol and the mitochondria. By tracing hydrogen in compartmentalized reactions that use NADPH as a cofactor, including the production of 2-hydroxyglutarate by mutant isocitrate dehydrogenase enzymes, we can observe metabolic pathway activity in these distinct cellular compartments. Using this system we determine the direction of serine/glycine interconversion within the mitochondria and cytosol, highlighting the ability of this approach to resolve compartmentalized reactions in intact cells.

journal_name

Mol Cell

journal_title

Molecular cell

authors

Lewis CA,Parker SJ,Fiske BP,McCloskey D,Gui DY,Green CR,Vokes NI,Feist AM,Vander Heiden MG,Metallo CM

doi

10.1016/j.molcel.2014.05.008

subject

Has Abstract

pub_date

2014-07-17 00:00:00

pages

253-63

issue

2

eissn

1097-2765

issn

1097-4164

pii

S1097-2765(14)00402-X

journal_volume

55

pub_type

杂志文章
  • Multiple mechanisms control chromosome integrity after replication fork uncoupling and restart at irreparable UV lesions.

    abstract::DNA replication forks pause in front of lesions on the template, eventually leading to cytotoxic chromosomal rearrangements. The in vivo structure of damaged eukaryotic replication intermediates has been so far elusive. Combining electron microscopy (EM) and two-dimensional (2D) gel electrophoresis, we found that UV-i...

    journal_title:Molecular cell

    pub_type: 杂志文章

    doi:10.1016/j.molcel.2005.11.015

    authors: Lopes M,Foiani M,Sogo JM

    更新日期:2006-01-06 00:00:00

  • Compartmentalization of the cell cortex by septins is required for maintenance of cell polarity in yeast.

    abstract::Formation and maintenance of specialized plasma membrane domains are crucial for many biological processes, such as cell polarization and signaling. During isotropic bud growth, the yeast cell periphery is divided into two domains: the bud surface, an active site of exocytosis and growth, and the relatively quiescent ...

    journal_title:Molecular cell

    pub_type: 杂志文章

    doi:10.1016/s1097-2765(00)80324-x

    authors: Barral Y,Mermall V,Mooseker MS,Snyder M

    更新日期:2000-05-01 00:00:00

  • Arginine methylation of FOXO transcription factors inhibits their phosphorylation by Akt.

    abstract::Forkhead box O (FOXO) transcription factors, the key regulators of cell survival, are negatively controlled through the PI3K-Akt signaling pathway. Phosphorylation of FOXO by Akt leads to cytoplasmic localization and subsequent degradation via the ubiquitin-proteasome system. Here we show a paradigm of FOXO1 regulatio...

    journal_title:Molecular cell

    pub_type: 杂志文章

    doi:10.1016/j.molcel.2008.09.013

    authors: Yamagata K,Daitoku H,Takahashi Y,Namiki K,Hisatake K,Kako K,Mukai H,Kasuya Y,Fukamizu A

    更新日期:2008-10-24 00:00:00

  • RNF8- and Ube2S-Dependent Ubiquitin Lysine 11-Linkage Modification in Response to DNA Damage.

    abstract::Ubiquitin modification of proteins plays pivotal roles in the cellular response to DNA damage. Given the complexity of ubiquitin conjugation due to the formation of poly-conjugates of different linkages, functional roles of linkage-specific ubiquitin modification at DNA damage sites are largely unclear. We identify th...

    journal_title:Molecular cell

    pub_type: 杂志文章

    doi:10.1016/j.molcel.2017.04.013

    authors: Paul A,Wang B

    更新日期:2017-05-18 00:00:00

  • Hit and run: transient deubiquitylase activity in a chromatin-remodeling complex.

    abstract::In this issue of Molecular Cell, Conaway and colleagues (Yao et al., 2008) provide a glimpse into an interesting mechanism to control a deubiquitylating enzyme via interaction of two complexes-the 19S proteasome regulatory particle and an ATP-dependent nucleosome remodeling complex. ...

    journal_title:Molecular cell

    pub_type: 评论,杂志文章

    doi:10.1016/j.molcel.2008.09.005

    authors: Zediak VP,Berger SL

    更新日期:2008-09-26 00:00:00

  • The 5'-7-methylguanosine cap on eukaryotic mRNAs serves both to stimulate canonical translation initiation and to block an alternative pathway.

    abstract::Translational control is frequently exerted at the stage of mRNA recruitment to the initiating ribosome. We have reconstituted mRNA recruitment to the 43S preinitiation complex (PIC) using purified S. cerevisiae components. We show that eIF3 and the eIF4 factors not only stabilize binding of mRNA to the PIC, they also...

    journal_title:Molecular cell

    pub_type: 杂志文章

    doi:10.1016/j.molcel.2010.08.021

    authors: Mitchell SF,Walker SE,Algire MA,Park EH,Hinnebusch AG,Lorsch JR

    更新日期:2010-09-24 00:00:00

  • Innate Immune Receptors as Competitive Determinants of Cell Fate.

    abstract::Infections can cause a multitude of stresses on the host and microbe. To detect potential infections, the mammalian immune system utilizes several families of pattern recognition receptors, which survey the intracellular and extracellular environments for microbial products. Members of each receptor family induce anti...

    journal_title:Molecular cell

    pub_type: 杂志文章,评审

    doi:10.1016/j.molcel.2017.05.009

    authors: Franz KM,Kagan JC

    更新日期:2017-06-15 00:00:00

  • PINK1 Phosphorylates MIC60/Mitofilin to Control Structural Plasticity of Mitochondrial Crista Junctions.

    abstract::Mitochondrial crista structure partitions vital cellular reactions and is precisely regulated by diverse cellular signals. Here, we show that, in Drosophila, mitochondrial cristae undergo dynamic remodeling among distinct subcellular regions and the Parkinson's disease (PD)-linked Ser/Thr kinase PINK1 participates in ...

    journal_title:Molecular cell

    pub_type: 杂志文章

    doi:10.1016/j.molcel.2018.01.026

    authors: Tsai PI,Lin CH,Hsieh CH,Papakyrikos AM,Kim MJ,Napolioni V,Schoor C,Couthouis J,Wu RM,Wszolek ZK,Winter D,Greicius MD,Ross OA,Wang X

    更新日期:2018-03-01 00:00:00

  • Acetylphosphate: a novel link between lysine acetylation and intermediary metabolism in bacteria.

    abstract::In this issue of Molecular Cell, Weinert et al. (2013) demonstrate that the intermediary metabolite acetyl-phosphate is an important acetyl donor that contributes to global protein acetylation in growth-arrested E. coli. ...

    journal_title:Molecular cell

    pub_type: 评论,杂志文章

    doi:10.1016/j.molcel.2013.07.006

    authors: Verdin E,Ott M

    更新日期:2013-07-25 00:00:00

  • ZNF410 Uniquely Activates the NuRD Component CHD4 to Silence Fetal Hemoglobin Expression.

    abstract::Metazoan transcription factors typically regulate large numbers of genes. Here we identify via a CRISPR-Cas9 genetic screen ZNF410, a pentadactyl DNA-binding protein that in human erythroid cells directly activates only a single gene, the NuRD component CHD4. Specificity is conveyed by two highly evolutionarily conser...

    journal_title:Molecular cell

    pub_type: 杂志文章

    doi:10.1016/j.molcel.2020.11.006

    authors: Lan X,Ren R,Feng R,Ly LC,Lan Y,Zhang Z,Aboreden N,Qin K,Horton JR,Grevet JD,Mayuranathan T,Abdulmalik O,Keller CA,Giardine B,Hardison RC,Crossley M,Weiss MJ,Cheng X,Shi J,Blobel GA

    更新日期:2021-01-21 00:00:00

  • Phosphatidic Acid Signals via the Hippo Pathway.

    abstract::In this issue of Molecular Cell, Han et al. (2018) show that phosphatidic acid inhibits the action of the LATS kinases, key mediators of Hippo pathway signaling, which leads to an increase in the nuclear activity of the transcriptional regulator YAP. ...

    journal_title:Molecular cell

    pub_type: 评论,杂志文章

    doi:10.1016/j.molcel.2018.10.004

    authors: Stampouloglou E,Varelas X

    更新日期:2018-10-18 00:00:00

  • The yeast nuclear cap binding complex can interact with translation factor eIF4G and mediate translation initiation.

    abstract::The mRNA cap structure is bound by either the nuclear (CBC) or the cytoplasmic (eIF4F) cap binding complex. Following mRNA export, CBC must be exchanged for eIF4F in the cytoplasm. It is not known how this exchange occurs or how this RNP remodeling event is integrated with mRNA function. Here we report genetic and bio...

    journal_title:Molecular cell

    pub_type: 杂志文章

    doi:

    authors: Fortes P,Inada T,Preiss T,Hentze MW,Mattaj IW,Sachs AB

    更新日期:2000-07-01 00:00:00

  • Codon Bias as a Means to Fine-Tune Gene Expression.

    abstract::The redundancy of the genetic code implies that most amino acids are encoded by multiple synonymous codons. In all domains of life, a biased frequency of synonymous codons is observed at the genome level, in functionally related genes (e.g., in operons), and within single genes. Other codon bias variants include biase...

    journal_title:Molecular cell

    pub_type: 杂志文章,评审

    doi:10.1016/j.molcel.2015.05.035

    authors: Quax TE,Claassens NJ,Söll D,van der Oost J

    更新日期:2015-07-16 00:00:00

  • Interaction of translation initiation factor 3 with the 30S ribosomal subunit.

    abstract::Hydroxyl radical footprinting and directed probing from Fe(II)-derivatized IF3 have been used to map the interaction of IF3 relative to 16S rRNA and tRNA(Met)(f) in the 30S ribosomal subunit. Our results place the two domains of IF3 on opposite sides of the initiator tRNA, with the C domain at the platform interface a...

    journal_title:Molecular cell

    pub_type: 杂志文章

    doi:10.1016/s1097-2765(01)00356-2

    authors: Dallas A,Noller HF

    更新日期:2001-10-01 00:00:00

  • Rictor forms a complex with Cullin-1 to promote SGK1 ubiquitination and destruction.

    abstract::The Rictor/mTOR complex (also known as mTORC2) plays a critical role in cellular homeostasis by phosphorylating AGC kinases such as Akt and SGK at their hydrophobic motifs to activate downstream signaling. However, the regulation of mTORC2 and whether it has additional function(s) remain largely unknown. Here, we repo...

    journal_title:Molecular cell

    pub_type: 杂志文章

    doi:10.1016/j.molcel.2010.08.016

    authors: Gao D,Wan L,Inuzuka H,Berg AH,Tseng A,Zhai B,Shaik S,Bennett E,Tron AE,Gasser JA,Lau A,Gygi SP,Harper JW,DeCaprio JA,Toker A,Wei W

    更新日期:2010-09-10 00:00:00

  • Viral-mediated noisy gene expression reveals biphasic E2f1 response to MYC.

    abstract::Gene expression mediated by viral vectors is subject to cell-to-cell variability, which limits the accuracy of gene delivery. When coupled with single-cell measurements, however, such variability provides an efficient means to quantify signaling dynamics in mammalian cells. Here, we illustrate the utility of this appr...

    journal_title:Molecular cell

    pub_type: 杂志文章

    doi:10.1016/j.molcel.2011.01.014

    authors: Wong JV,Yao G,Nevins JR,You L

    更新日期:2011-02-04 00:00:00

  • Mechanistic Insights into the cis- and trans-Acting DNase Activities of Cas12a.

    abstract::CRISPR-Cas12a (Cpf1) is an RNA-guided DNA-cutting nuclease that has been repurposed for genome editing. Upon target DNA binding, Cas12a cleaves both the target DNA in cis and non-target single-stranded DNAs (ssDNAs) in trans. To elucidate the molecular basis for both DNase cleavage modes, we performed structural and b...

    journal_title:Molecular cell

    pub_type: 杂志文章

    doi:10.1016/j.molcel.2018.11.021

    authors: Swarts DC,Jinek M

    更新日期:2019-02-07 00:00:00

  • EGFR-Phosphorylated Platelet Isoform of Phosphofructokinase 1 Promotes PI3K Activation.

    abstract::EGFR activates phosphatidylinositide 3-kinase (PI3K), but the mechanism underlying this activation is not completely understood. We demonstrated here that EGFR activation resulted in lysine acetyltransferase 5 (KAT5)-mediated K395 acetylation of the platelet isoform of phosphofructokinase 1 (PFKP) and subsequent trans...

    journal_title:Molecular cell

    pub_type: 杂志文章

    doi:10.1016/j.molcel.2018.03.018

    authors: Lee JH,Liu R,Li J,Wang Y,Tan L,Li XJ,Qian X,Zhang C,Xia Y,Xu D,Guo W,Ding Z,Du L,Zheng Y,Chen Q,Lorenzi PL,Mills GB,Jiang T,Lu Z

    更新日期:2018-04-19 00:00:00

  • Dynamics of substrate denaturation and translocation by the ClpXP degradation machine.

    abstract::ClpXP is a protein machine composed of the ClpX ATPase, a member of the Clp/Hsp100 family of remodeling enzymes, and the ClpP peptidase. Here, ClpX and ClpXP are shown to catalyze denaturation of GFP modified with an ssrA degradation tag. ClpX translocates this denatured protein into the proteolytic chamber of ClpP an...

    journal_title:Molecular cell

    pub_type: 杂志文章

    doi:10.1016/s1097-2765(00)80243-9

    authors: Kim YI,Burton RE,Burton BM,Sauer RT,Baker TA

    更新日期:2000-04-01 00:00:00

  • Introns Protect Eukaryotic Genomes from Transcription-Associated Genetic Instability.

    abstract::Transcription is a source of genetic instability that can notably result from the formation of genotoxic DNA:RNA hybrids, or R-loops, between the nascent mRNA and its template. Here we report an unexpected function for introns in counteracting R-loop accumulation in eukaryotic genomes. Deletion of endogenous introns i...

    journal_title:Molecular cell

    pub_type: 杂志文章

    doi:10.1016/j.molcel.2017.07.002

    authors: Bonnet A,Grosso AR,Elkaoutari A,Coleno E,Presle A,Sridhara SC,Janbon G,Géli V,de Almeida SF,Palancade B

    更新日期:2017-08-17 00:00:00

  • Crystal structure and deletion analysis show that the accessory subunit of mammalian DNA polymerase gamma, Pol gamma B, functions as a homodimer.

    abstract::Polymerase gamma, which replicates and repairs mitochondrial DNA, requires the Pol gamma B subunit for processivity. We determined the crystal structure of mouse Pol gamma B, a core component of the mitochondrial replication machinery. Pol gamma B shows high similarity to glycyl-tRNA synthetase and dimerizes through a...

    journal_title:Molecular cell

    pub_type: 杂志文章

    doi:10.1016/s1097-2765(01)00153-8

    authors: Carrodeguas JA,Theis K,Bogenhagen DF,Kisker C

    更新日期:2001-01-01 00:00:00

  • Structure of the papillomavirus DNA-tethering complex E2:Brd4 and a peptide that ablates HPV chromosomal association.

    abstract::Many DNA viruses that are latent in dividing cells are noncovalent passengers on mitotic chromosomes and require specific viral-encoded and cellular factors for this activity. The chromosomal protein Brd4 is implicated in the hitchhiking of bovine papillomavirus-1 (BPV-1), and the viral protein E2 binds to both plasmi...

    journal_title:Molecular cell

    pub_type: 杂志文章

    doi:10.1016/j.molcel.2006.11.002

    authors: Abbate EA,Voitenleitner C,Botchan MR

    更新日期:2006-12-28 00:00:00

  • A bitter PP1 fights the sweet polo.

    abstract::In a recent paper in Developmental Cell, Yamashiro et al. (2008) report that the PP1 regulatory subunit MYPT1 interacts with PLK1 and antagonizes essential mitotic functions of PLK1, at least in part by promoting the dephosphorylation of PLK1 at Thr210. ...

    journal_title:Molecular cell

    pub_type: 杂志文章

    doi:10.1016/j.molcel.2008.05.012

    authors: Archambault V,Chen F,Glover DM

    更新日期:2008-06-06 00:00:00

  • The K48-K63 Branched Ubiquitin Chain Regulates NF-κB Signaling.

    abstract::Polyubiquitin chains of different topologies regulate diverse cellular processes. K48- and K63-linked chains, the two most abundant chain types, regulate proteolytic and signaling pathways, respectively. Although recent studies reported important roles for heterogeneous chains, the functions of branched ubiquitin chai...

    journal_title:Molecular cell

    pub_type: 杂志文章

    doi:10.1016/j.molcel.2016.09.014

    authors: Ohtake F,Saeki Y,Ishido S,Kanno J,Tanaka K

    更新日期:2016-10-20 00:00:00

  • High-Resolution Mapping of Multiway Enhancer-Promoter Interactions Regulating Pathogen Detection.

    abstract::Eukaryotic gene expression regulation involves thousands of distal regulatory elements. Understanding the quantitative contribution of individual enhancers to gene expression is critical for assessing the role of disease-associated genetic risk variants. Yet, we lack the ability to accurately link genes with their dis...

    journal_title:Molecular cell

    pub_type: 杂志文章

    doi:10.1016/j.molcel.2020.09.005

    authors: Vangala P,Murphy R,Quinodoz SA,Gellatly K,McDonel P,Guttman M,Garber M

    更新日期:2020-10-15 00:00:00

  • WTX: an unexpected regulator for p53.

    abstract::The WTX gene is frequently lost or mutated in Wilms tumor. In this issue of Molecular Cell, Kim et al. (2012) identify WTX modulation of p53 tumor-suppressor activity through regulation of p53 acetylation. Therefore, WTX differentially regulates the oncogenic β-catenin pathway and the tumor-suppressing p53 pathway. ...

    journal_title:Molecular cell

    pub_type: 评论,杂志文章

    doi:10.1016/j.molcel.2012.02.010

    authors: Dai C,Gu W

    更新日期:2012-03-09 00:00:00

  • Mir-214-dependent regulation of the polycomb protein Ezh2 in skeletal muscle and embryonic stem cells.

    abstract::Polycomb group (PcG) proteins exert essential functions in the most disparate biological processes. The contribution of PcG proteins to cell commitment and differentiation relates to their ability to repress transcription of developmental regulators in embryonic stem (ES) cells and in committed cell lineages, includin...

    journal_title:Molecular cell

    pub_type: 杂志文章

    doi:10.1016/j.molcel.2009.08.008

    authors: Juan AH,Kumar RM,Marx JG,Young RA,Sartorelli V

    更新日期:2009-10-09 00:00:00

  • Differential targeting of prosurvival Bcl-2 proteins by their BH3-only ligands allows complementary apoptotic function.

    abstract::Apoptosis is initiated when Bcl-2 and its prosurvival relatives are engaged by proapoptotic BH3-only proteins via interaction of its BH3 domain with a groove on the Bcl-2-like proteins. These interactions have been considered promiscuous, but our analysis of the affinity of eight BH3 peptides for five Bcl-2-like prote...

    journal_title:Molecular cell

    pub_type: 杂志文章

    doi:10.1016/j.molcel.2004.12.030

    authors: Chen L,Willis SN,Wei A,Smith BJ,Fletcher JI,Hinds MG,Colman PM,Day CL,Adams JM,Huang DC

    更新日期:2005-02-04 00:00:00

  • The little elongation complex functions at initiation and elongation phases of snRNA gene transcription.

    abstract::The small nuclear RNA (snRNA) genes have been widely used as a model system for understanding transcriptional regulation due to the unique aspects of their promoter structure, selectivity for either RNA polymerase (Pol) II or III, and because of their unique mechanism of termination that is tightly linked with the pro...

    journal_title:Molecular cell

    pub_type: 杂志文章

    doi:10.1016/j.molcel.2013.07.003

    authors: Hu D,Smith ER,Garruss AS,Mohaghegh N,Varberg JM,Lin C,Jackson J,Gao X,Saraf A,Florens L,Washburn MP,Eissenberg JC,Shilatifard A

    更新日期:2013-08-22 00:00:00

  • Gemin5 delivers snRNA precursors to the SMN complex for snRNP biogenesis.

    abstract::The SMN complex assembles Sm cores on snRNAs, a key step in the biogenesis of snRNPs, the spliceosome's major components. Here, using SMN complex inhibitors identified by high-throughput screening and a ribo-proteomic strategy on formaldehyde crosslinked RNPs, we dissected this pathway in cells. We show that protein s...

    journal_title:Molecular cell

    pub_type: 杂志文章

    doi:10.1016/j.molcel.2010.03.014

    authors: Yong J,Kasim M,Bachorik JL,Wan L,Dreyfuss G

    更新日期:2010-05-28 00:00:00