Deletion Strains Reveal Metabolic Roles for Key Elemental Sulfur-Responsive Proteins in Pyrococcus furiosus

被引:47
作者
Bridger, Stephanie L. [1 ]
Clarkson, Sonya M. [1 ]
Stirrett, Karen [2 ]
DeBarry, Megan B. [2 ]
Lipscomb, Gina L. [2 ]
Schut, Gerrit J. [1 ]
Westpheling, Janet [2 ]
Scott, Robert A. [1 ]
Adams, Michael W. W. [1 ]
机构
[1] Univ Georgia, Dept Biochem & Mol Biol, Athens, GA 30602 USA
[2] Univ Georgia, Dept Genet, Athens, GA 30602 USA
关键词
DNA MICROARRAY ANALYSIS; THERMOCOCCUS-KODAKARENSIS; SULFIDE DEHYDROGENASE; HYDROGEN-PRODUCTION; ARCHAEON; HYPERTHERMOPHILE; OXIDOREDUCTASE; FLAVOPROTEIN; REDUCTION; ENZYMES;
D O I
10.1128/JB.05445-11
中图分类号
Q93 [微生物学];
学科分类号
071005 ; 100705 ;
摘要
Transcriptional and enzymatic analyses of Pyrococcus furiosus previously indicated that three proteins play key roles in the metabolism of elemental sulfur (S-0): a membrane-bound oxidoreductase complex (MBX), a cytoplasmic coenzyme A-dependent NADPH sulfur oxidoreductase (NSR), and sulfur-induced protein A (SipA). Deletion strains, referred to as MBX1, NSR1, and SIP1, respectively, have now been constructed by homologous recombination utilizing the uracil auxotrophic COM1 parent strain (Delta pyrF). The growth of all three mutants on maltose was comparable without S-0, but in its presence, the growth of MBX1 was greatly impaired while the growth of NSR1 and SIP1 was largely unaffected. In the presence of S-0, MBX1 produced little, if any, sulfide but much more acetate (per unit of protein) than the parent strain, demonstrating that MBX plays a critical role in S-0 reduction and energy conservation. In contrast, comparable amounts of sulfide and acetate were produced by NSR1 and the parent strain, indicating that NSR is not essential for energy conservation during S-0 reduction. Differences in transcriptional responses to S-0 in NSR1 suggest that two sulfide dehydrogenase isoenzymes provide a compensatory NADPH-dependent S-0 reduction system. Genes controlled by the S-0-responsive regulator SurR were not as highly regulated in MBX1 and NSR1. SIP1 produced the same amount of acetate but more sulfide than the parent strain. That SipA is not essential for growth on S-0 indicates that it is not required for detoxification of metal sulfides, as previously suggested. A model is proposed for S-0 reduction by P. furiosus with roles for MBX and NSR in bioenergetics and for SipA in iron-sulfur metabolism.
引用
收藏
页码:6498 / 6504
页数:7
相关论文
共 27 条
[1]   Key role for sulfur in peptide metabolism and in regulation of three hydrogenases in the hyperthermophilic archaeon Pyrococcus furiosus [J].
Adams, MWW ;
Holden, JF ;
Menon, AL ;
Schut, GJ ;
Grunden, AM ;
Hou, C ;
Hutchins, AM ;
Jenney, FE ;
Kim, C ;
Ma, KS ;
Pan, GL ;
Roy, R ;
Sapra, R ;
Story, SV ;
Verhagen, MFJM .
JOURNAL OF BACTERIOLOGY, 2001, 183 (02) :716-724
[2]  
BRADFORD MM, 1976, ANAL BIOCHEM, V72, P248, DOI 10.1016/0003-2697(76)90527-3
[3]   INHIBITION OF METHYLENE-BLUE FORMATION DURING DETERMINATION OF ACID-LABILE SULFIDE OF IRON-SULFUR PROTEIN SAMPLES CONTAINING DITHIONITE [J].
CHEN, JS ;
MORTENSON, LE .
ANALYTICAL BIOCHEMISTRY, 1977, 79 (1-2) :157-165
[4]   The Elemental Sulfur-Responsive Protein (SipA) from the Hyperthermophilic Archaeon Pyrococcus furiosus Is Regulated by Sulfide in an Iron-Dependent Manner [J].
Clarkson, Sonya M. ;
Newcomer, Elizabeth C. ;
Young, Everett G. ;
Adams, Michael W. W. .
JOURNAL OF BACTERIOLOGY, 2010, 192 (21) :5841-5843
[5]   PYROCOCCUS-FURIOSUS SP-NOV REPRESENTS A NOVEL GENUS OF MARINE HETEROTROPHIC ARCHAEBACTERIA GROWING OPTIMALLY AT 100-DEGREES C [J].
FIALA, G ;
STETTER, KO .
ARCHIVES OF MICROBIOLOGY, 1986, 145 (01) :56-61
[6]   Novel structure and redox chemistry of the prosthetic groups of the iron-sulfur flavoprotein sulfide dehydrogenase from Pyrococcus furiosus;: evidence for a [2Fe-2S] cluster with Asp(Cys)3 ligands [J].
Hagen, WR ;
Silva, PJ ;
Amorim, MA ;
Hagedoorn, PL ;
Wassink, H ;
Haaker, H ;
Robb, FT .
JOURNAL OF BIOLOGICAL INORGANIC CHEMISTRY, 2000, 5 (04) :527-534
[7]   Nucleic acids bind to nanoparticulate iron (II) monosulphide in aqueous solutions [J].
Hatton, Bryan ;
Rickard, David .
ORIGINS OF LIFE AND EVOLUTION OF BIOSPHERES, 2008, 38 (03) :257-270
[8]   Micromolar intracellular hydrogen peroxide disrupts metabolism by damaging iron-sulfur enzymes [J].
Jang, Soojin ;
Imlay, James A. .
JOURNAL OF BIOLOGICAL CHEMISTRY, 2007, 282 (02) :929-937
[9]   Distinct Physiological Roles of the Three [NiFe]-Hydrogenase Orthologs in the Hyperthermophilic Archaeon Thermococcus kodakarensis [J].
Kanai, Tamotsu ;
Matsuoka, Ryoji ;
Beppu, Haruki ;
Nakajima, Akihito ;
Okada, Yoshihiro ;
Atomi, Haruyuki ;
Imanaka, Tadayuki .
JOURNAL OF BACTERIOLOGY, 2011, 193 (12) :3109-3116
[10]   Kinetics of the chemical oxidation of polysulfide anions in aqueous solution [J].
Kleinjan, WE ;
de Keizer, A ;
Janssen, AJH .
WATER RESEARCH, 2005, 39 (17) :4093-4100