Cloning and characterization of the thiD/J gene of Escherichia coli encoding a thiamin-synthesizing bifunctional enzyme, hydroxymethylpyrimidine kinase/phosphomethylpyrimidine kinase

Mizote, Tomoko; Tsuda, Masashi; Smith, D. G.; Nakayama, Hideo; Nakazawa, Takahiro

doi:10.1099/13500872-145-2-495

Cited by 50 publications

(34 citation statements)

References 24 publications

Supporting

Mentioning

Contrasting

Unclassified

Order By: Relevance

“…ThiJ was a thiamin biosynthesis enzyme that catalyzed the phosphorylation of hydroxymethylpyrimidine (HMP) to HMP monophosphate. The enzyme was primarily studied in E. coli (Mizote et al, 1999), but has not been characterized in plants. There are two other Arabidopsis proteins that are highly homologous to the cabbage protein (BLASTP value E < 10 −70…”

Section: Resultsmentioning

confidence: 99%

Molecular Characterization of a thiJ-like Gene in Chinese Cabbage

Park²,

Lee³

et al. 2004

BMB Reports

View full text Add to dashboard Cite

A cDNA clone for a salicylic acid-induced gene in Chinese cabbage (Brassica rapa subsp. pekinensis) was isolated and characterized. The cabbage gene encoding a protein of 392 amino acids contained a tandem array of two thiJ-like sequences. ThiJ is a thiamin biosynthesis enzyme that catalyzes the phosphorylation of hydroxymethylpyrimidine (HMP) to HMP monophosphate. Although the cabbage gene shows a similarity to bacterial thiJ genes, it also shares a similarity with the human DJ-1, a multifunctional protein that is involved in transcription regulation, male fertility, and parkinsonism. The cabbage thiJ-like gene is strongly induced by salicylic acid and a nonhost pathogen, Pseudomonas syringae pv. tomato, which elicits a hypersensitive response in Chinese cabbage. Treatment of the cabbage leaves with BTH, methyl jasmonate, or ethephon showed that the cabbage thiJ-like gene expression is also strongly induced by BTH, but not by methyl jasmonate or ethylene. This indicates that the cabbage gene is activated via a salicylic acid-dependent signaling pathway. Examination of the tissue-specific expression revealed that the induction of the cabbage gene expression by BTH occurs in the leaf, stem, and floral tissues but not in the root.

show abstract

Section: Resultsmentioning

confidence: 99%

Molecular Characterization of a thiJ-like Gene in Chinese Cabbage

Park²,

Lee³

et al. 2004

BMB Reports

View full text Add to dashboard Cite

show abstract

“…However, in vitro studies have shown that HMP itself does not bind to the riboswitch (27), and our own studies here indicate that neither HMP nor HMP-P competes with HMP-PP binding. If we therefore assume that this also applies in vivo, then it is highly plausible that HMP is pyrophosphorylated by THID (32) and can bind to the riboswitch in a similar manner to TPP. Direct evidence that HMP-PP binds to the THIC riboswitch is provided by equilibrium dialysis (Fig.…”

Section: Discussionmentioning

confidence: 99%

Analysis of Chlamydomonas thiamin metabolism in vivo reveals riboswitch plasticity

Moulin

Nguyen

Scaife

et al. 2013

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

View full text Add to dashboard Cite

Thiamin (vitamin B 1 ) is an essential micronutrient needed as a cofactor for many central metabolic enzymes. Animals must have thiamin in their diet, whereas bacteria, fungi, and plants can biosynthesize it de novo from the condensation of a thiazole and a pyrimidine moiety. Although the routes to biosynthesize these two heterocycles are not conserved in different organisms, in all cases exogenous thiamin represses expression of one or more of the biosynthetic pathway genes. One important mechanism for this control is via thiamin-pyrophosphate (TPP) riboswitches, regions of the mRNA to which TPP can bind directly, thus facilitating fine-tuning to maintain homeostasis. However, there is little information on how modulation of riboswitches affects thiamin metabolism in vivo. Here we use the green alga, Chlamydomonas reinhardtii, which regulates both thiazole and pyrimidine biosynthesis with riboswitches in the THI4 (Thiamin 4) and THIC (Thiamin C) genes, respectively, to investigate this question. Our study reveals that regulation of thiamin metabolism is not the simple dogma of negative feedback control. Specifically, balancing the provision of both of the heterocycles of TPP appears to be an important requirement. Furthermore, we show that the Chlamydomonas THIC riboswitch is controlled by hydroxymethylpyrimidine pyrophosphate, as well as TPP, but with an identical alternative splicing mechanism. Similarly, the THI4 gene is responsive to thiazole. The study not only provides insight into the plasticity of the TPP riboswitches but also shows that their maintenance is likely to be a consequence of evolutionary need as a function of the organisms' environment and the particular pathway used.eukaryotic riboswitch | gene expression coordination | metabolic regulation | cross-talk

show abstract

“…This family includes proteins involved in RNA-protein interaction regulation, thiamine biosynthesis, and protease activity, which do not appear to be related to one another. The role of this family remains unclear, and most of the members are predicted proteins that have, to our knowledge, not been functionally analyzed except for ThiJ, which catalyzes hydroxymethylpyrimidine phosphorylation in the thiamine biosynthetic pathway (35,36), and for intracellular proteases (PfpI and PH1704) in hyperthermophilic archaea (30,31). Here, we discovered that the members of the ThiJ/PfpI family and isonitrile hydratase (which is an enzyme playing a novel physiological role in isonitrile degradation) comprise a novel superfamily.…”

Section: Fig 4 CD Spectra Of the Wild-type And Mutant Isonitrile Hymentioning

confidence: 99%

Isonitrile Hydratase from Pseudomonas putidaN19–2

Goda

Hashimoto

Takase

et al. 2002

Journal of Biological Chemistry

View full text Add to dashboard Cite

Isonitrile hydratase is a novel enzyme in Pseudomonas putida N19 -2 that catalyzes the conversion of isonitriles to N-substituted formamides. Based on N-terminal and internal amino acid sequences, a 535-bp DNA fragment corresponding to a portion of the isonitrile hydratase gene was amplified, which was used as a probe to clone a 6.4-kb DNA fragment containing the whole gene. Sequence analysis of the 6.4-kb fragment revealed that the isonitrile hydratase gene (inhA) was 684 nucleotides long and encoded a protein with a molecular mass of 24,211 Da. Overexpression of inhA in Escherichia coli gave a large amount of soluble isonitrile hydratase exhibiting the same molecular and catalytic properties as the native enzyme from the Pseudomonas strain. The predicted amino acid sequence of inhA showed low similarity to that of an intracellular protease in Pyrococcus horikoshii (PH1704), and an active cysteine residue in the protease was conserved in the isonitrile hydratase at the corresponding position (Cys-101). A mutant enzyme containing Ala instead of Cys-101 did not exhibit isonitrile hydratase activity at all, demonstrating the essential role of this residue in the catalytic function.

show abstract

Cloning and characterization of the thiD/J gene of Escherichia coli encoding a thiamin-synthesizing bifunctional enzyme, hydroxymethylpyrimidine kinase/phosphomethylpyrimidine kinase

Cited by 50 publications

References 24 publications

Molecular Characterization of a thiJ-like Gene in Chinese Cabbage

Molecular Characterization of a thiJ-like Gene in Chinese Cabbage

Analysis of Chlamydomonas thiamin metabolism in vivo reveals riboswitch plasticity

Isonitrile Hydratase from Pseudomonas putidaN19–2

Contact Info

Product

Resources

About