Posttranslational synthesis of hypusine: evolutionary progression and specificity of the hypusine modification

Wolff, Edith C.; Kang, Kee Ryeon; Kim, So Hyun; Park, M. H.

doi:10.1007/s00726-007-0525-0

Cited by 128 publications

(119 citation statements)

References 49 publications

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“…Of course, the growth experiments reported in the present study do not exclude other important physiological functions for polyamines that are not reflected in growth of E. coli or the physiological functions of polyamines in other organisms. In yeast and other eukaryotes, for example, spermidine is clearly required for growth because of its conversion to hypusine in the eukaryote protein synthesis factor eIF-5A (3,26).…”

Section: Discussionmentioning

confidence: 99%

Polyamines Are Not Required for Aerobic Growth of Escherichia coli : Preparation of a Strain with Deletions in All of the Genes for Polyamine Biosynthesis

Chattopadhyay

Tabor

2009

J Bacteriol

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A strain of Escherichia coli was constructed in which all of the genes involved in polyamine biosynthesisspeA (arginine decarboxylase), speB (agmatine ureohydrolase), speC (ornithine decarboxylase), spe D (adenosylmethionine decarboxylase), speE (spermidine synthase), speF (inducible ornithine decarboxylase), cadA (lysine decarboxylase), and ldcC (lysine decarboxylase)-had been deleted. Despite the complete absence of all of the polyamines, the strain grew indefinitely in air in amine-free medium, albeit at a slightly (ca. 40 to 50%) reduced growth rate. Even though this strain grew well in the absence of the amines in air, it was still sensitive to oxygen stress in the absence of added spermidine. In contrast to the ability to grow in air in the absence of polyamines, this strain, surprisingly, showed a requirement for polyamines for growth under strictly anaerobic conditions.Polyamines are highly abundant in essentially all organisms, ranging from bacteria to humans, and there have been a large number of studies from this and many other laboratories reporting a variety of phenotypic effects resulting from changes in the concentration of polyamines in both in vitro and in vivo experiments. In particular, polyamines have been associated with such biological processes as nucleic acid and protein biosynthesis and structure, cell growth, and differentiation (reviewed in references 5, 22, and 23). Therefore, it was surprising that, in our earlier studies (24), we found that a mutant of Escherichia coli that had mutations in the genes for the biosynthesis of the polyamines (⌬speA, ⌬speB, ⌬speC, ⌬speD, ⌬speE, and cadA) still grew indefinitely in a polyamine-free medium, albeit at a decreased growth rate (ca. 30% of the normal growth rate).The strain used in our previous studies still had trace amounts of putrescine and significant amounts of cadaverine. To study whether these small amounts of amines could account for the slow growth of these strains, we have now constructed a new strain that is completely deficient in these amines by including deletions of cadA (inducible lysine decarboxylase), ldcC (constitutive lysine decarboxylase), and speF (inducible ornithine decarboxylase) to the strain described above. We found that this strain which is completely deficient in all of the amines still grows well (40 to 50% of normal growth rate) in purified medium in air. This indicates that, at least for this organism, the various physiological functions attributed to polyamines are not required for growth in air. In contrast, we have found that polyamines are required for growth of this strain in 95% oxygen and under anaerobic conditions. MATERIALS AND METHODSStrain construction. P1 transductions were carried out essentially as described by Miller (17). The strains used for the P1 transduction were obtained from the Keio collection (1) from the Yale E. coli Genetics Center. The deleted genes in this collection contain a kanamycin insert, which was used for the selection process. For use in subsequent transduction experiments,...

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Section: Discussionmentioning

confidence: 99%

Polyamines Are Not Required for Aerobic Growth of Escherichia coli : Preparation of a Strain with Deletions in All of the Genes for Polyamine Biosynthesis

Chattopadhyay

Tabor

2009

J Bacteriol

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“…eIF5A is post-translationally modified by the conversion of a lysine residue to hypusine (Wolff et al 2007;Dever et al 2014). An n-butylamine group is transferred from spermidine to the e-amino group of Lys51 in eIF5A to generate deoxyhypusine.…”

Section: Eif5a Promotion Of Peptide Bond Formationmentioning

confidence: 99%

Mechanism and Regulation of Protein Synthesis in Saccharomyces cerevisiae

2016

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In this review, we provide an overview of protein synthesis in the yeast Saccharomyces cerevisiae. The mechanism of protein synthesis is well conserved between yeast and other eukaryotes, and molecular genetic studies in budding yeast have provided critical insights into the fundamental process of translation as well as its regulation. The review focuses on the initiation and elongation phases of protein synthesis with descriptions of the roles of translation initiation and elongation factors that assist the ribosome in binding the messenger RNA (mRNA), selecting the start codon, and synthesizing the polypeptide. We also examine mechanisms of translational control highlighting the mRNA cap-binding proteins and the regulation of GCN4 and CPA1 mRNAs.

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“…H ypusine is an unusual, but highly conserved, amino acid that is found only in the eukaryotic translational initiation factor 5A (eIF5A), a protein that regulates cell proliferation (1,2). The biosynthesis of eIF5A involves a posttranslational modification of the eIF5A precursor, where a lysine residue is first modified to deoxyhypusine (Dhp) by deoxyhypusine synthase (DHS) and then the nascent Dhp is hydroxylated by deoxyhypusine hydroxylase (DOHH) to form hypusine (Hpu) (Scheme 1) (1, 2).…”

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confidence: 99%

Human deoxyhypusine hydroxylase, an enzyme involved in regulating cell growth, activates O ₂ with a nonheme diiron center

Emerson

Martinho

et al. 2009

Proc. Natl. Acad. Sci. U.S.A.

107

141

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Deoxyhypusine hydroxylase is the key enzyme in the biosynthesis of hypusine containing eukaryotic translation initiation factor 5A (eIF5A), which plays an essential role in the regulation of cell proliferation. Recombinant human deoxyhypusine hydroxylase (hDOHH) has been reported to have oxygen-and iron-dependent activity, an estimated iron/holoprotein stoichiometry of 2, and a visible band at 630 nm responsible for the blue color of the as-isolated protein. EPR, Mö ssbauer, and XAS spectroscopic results presented herein provide direct spectroscopic evidence that hDOHH has an antiferromagnetically coupled diiron center with histidines and carboxylates as likely ligands, as suggested by mutagenesis experiments. Resonance Raman experiments show that its blue chromophore arises from a ( -1,2-peroxo)diiron(III) center that forms in the reaction of the reduced enzyme with O 2, so the peroxo form of hDOHH is unusually stable. Nevertheless we demonstrate that it can carry out the hydroxylation of the deoxyhypusine residue present in the elF5A substrate. Despite a lack of sequence similarity, hDOHH has a nonheme diiron active site that resembles both in structure and function those found in methane and toluene monooxygenases, bacterial and mammalian ribonucleotide reductases, and stearoyl acyl carrier protein ⌬ 9 -desaturase from plants, suggesting that the oxygen-activating diiron motif is a solution arrived at by convergent evolution. Notably, hDOHH is the only example thus far of a human hydroxylase with such a diiron active site.H ypusine is an unusual, but highly conserved, amino acid that is found only in the eukaryotic translational initiation factor 5A (eIF5A), a protein that regulates cell proliferation (1, 2). The biosynthesis of eIF5A involves a posttranslational modification of the eIF5A precursor, where a lysine residue is first modified to deoxyhypusine (Dhp) by deoxyhypusine synthase (DHS) and then the nascent Dhp is hydroxylated by deoxyhypusine hydroxylase (DOHH) to form hypusine (Hpu) (Scheme 1) (1, 2). The importance of hypusine and these 2 enzymes has been shown by several studies where depletion of spermidine (3) or inhibition of either DHS or DOHH (4, 5) leads to a decrease of hypusinecontaining eIF5A [eIF5A(Hpu)] and inhibition of eukaryotic cell growth. Consequently, these results suggest that eIF5A and DOHH could be promising targets for antitumor (6) and anti-HIV-1 therapies (7).The hydroxylase activity of recombinant human DOHH (hDOHH) has been shown to depend on Fe(II) and not on any other physiologically relevant divalent metal ion. An estimated iron-to-holoprotein stoichiometry of 2 is observed (8). Sequence examination, homology modeling, and mutagenesis experiments suggest 2 possible iron binding sites consisting of histidine and carboxylate ligands (8,9). Thus at first glance, hDOHH appears to resemble members of the superfamily of bacterial diiron multicomponent monooxygenases, like methane or toluene monooxygenase, that use nonheme diiron centers to activate dioxygen for the hydroxylat...

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Posttranslational synthesis of hypusine: evolutionary progression and specificity of the hypusine modification

Cited by 128 publications

References 49 publications

Polyamines Are Not Required for Aerobic Growth of Escherichia coli : Preparation of a Strain with Deletions in All of the Genes for Polyamine Biosynthesis

Polyamines Are Not Required for Aerobic Growth of Escherichia coli : Preparation of a Strain with Deletions in All of the Genes for Polyamine Biosynthesis

Mechanism and Regulation of Protein Synthesis in Saccharomyces cerevisiae

Human deoxyhypusine hydroxylase, an enzyme involved in regulating cell growth, activates O ₂ with a nonheme diiron center

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Posttranslational synthesis of hypusine: evolutionary progression and specificity of the hypusine modification

Cited by 128 publications

References 49 publications

Polyamines Are Not Required for Aerobic Growth of Escherichia coli : Preparation of a Strain with Deletions in All of the Genes for Polyamine Biosynthesis

Polyamines Are Not Required for Aerobic Growth of Escherichia coli : Preparation of a Strain with Deletions in All of the Genes for Polyamine Biosynthesis

Mechanism and Regulation of Protein Synthesis in Saccharomyces cerevisiae

Human deoxyhypusine hydroxylase, an enzyme involved in regulating cell growth, activates O 2 with a nonheme diiron center

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Product

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Human deoxyhypusine hydroxylase, an enzyme involved in regulating cell growth, activates O ₂ with a nonheme diiron center