Cloning and characteristics of a positive regulatory gene, THI2 (PHO6), of thiamin biosynthesis in Saccharomyces cerevisiae

Nishimura, Hiroshi; Kawasaki, Yuko; Kaneko, Yoshinobu; Nosaka, Kazuto; Iwashima, Akio

doi:10.1016/0014-5793(92)80349-l

Cited by 49 publications

(31 citation statements)

References 11 publications

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“…Similar results were obtained with the yeast S. cerevisiae (11)(12)(13). Two regulatory genes are involved in the expression of the thiamine-sensitive genes.…”

Section: Transportsupporting

confidence: 84%

Regulation of Gene Expression by Cofactors Derived from B Vitamins.

Brandsch

1994

Journal of Nutritional Science and Vitaminology, J Nutr Sci Vit

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SummaryVitamins represent precursor molecules for the majority of cofactors essential for various enzyme activities. Is the expression of proteins that require a specific cofactor for their function correlated to its availability? Examples are presented and discussed of the involvement of vitamin-derived cofactors in the regulatory mechanisms controlling gene expression in bacteria and eukaryotes at the transcriptional and trans lational level.

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“…Similar results were obtained with the yeast S. cerevisiae (11)(12)(13). Two regulatory genes are involved in the expression of the thiamine-sensitive genes.…”

Section: Transportsupporting

confidence: 84%

Regulation of Gene Expression by Cofactors Derived from B Vitamins.

Brandsch

1994

Journal of Nutritional Science and Vitaminology, J Nutr Sci Vit

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“…Another thiamine auxotrophic mutant carrying thi2 (pho6) (9) also showed decreased T-rAPase activity, whereas thiamine transport activity was retained in this mutant (16). A complementation test between the thi2 (pho6) mutant (K269-Sc) and the thi3 mutant (TRS3) isolated in this study was then carried out.…”

Section: Methodsmentioning

confidence: 94%

“…We have already elucidated that expression of the structural genes for T-rAPase and the enzymes involved in thiamine biosynthesis in S. cerevisiae is regulated positively by THI2 (PH06) (9), whereas expression of the genes for thiamine metabolism is controlled negatively by the intracellular thiamine pyrophosphate level (15). However, as described above, the THI2 (PH06) gene seems not to be essential for expression of the thiamine transport system (16), and therefore we have attempted to obtain a thiamine auxotrophic mutant with a defect in the thiamine transport system in order to identify a positive regulatory gene for thiamine metabolism, including thiamine transport. As expected, the mutant (TRS3) isolated in this study was auxotrophic for thiamine, with defective T-rAPase and thiamine transport activities.…”

Section: Bofmentioning

confidence: 96%

“…Since PHO6 is closely related to thiamine metabolism, we described this gene as THI2 (PHO6). However, the thi2 (pho6) mutant defective in the regulatory gene for enzymes involved in thiamine synthesis retained sufficient thiamine transport activity (16). Another thiamine auxotrophic strain (thil mutant) was reported (5), and it was found that the allelic thil locus is located in chromosome XIII (14).…”

mentioning

confidence: 99%

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A positive regulatory gene, THI3, is required for thiamine metabolism in Saccharomyces cerevisiae

et al. 1992

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We have isolated a thiamine auxotrophic mutant carrying a recessive mutation which lacks the positive regulatory gene, THI3, which differs in the regulation of thiamine transport from the THI2 (PHO6) gene described previously (Y. Kawasaki, K. Nosaka, Y. Kaneko, H. Nishimura, and A. Iwashima, J. Bacteriol. 172:6145-6147, 1990) for expression of thiamine metabolism in Saccharomyces cerevisiae. The mutant (thi3) had a markedly reduced thiamine transport system as well as reduced activity of thiamine-repressible acid phosphatase and of several enzymes for thiamine synthesis from 2-methyl-4-amino-5-hydroxymethylpyrimidine and 4-methyl-5-1-hydroxyethylthiazole. These results suggest that thiamine metabolism in S. cerevisiae is subject to two positive regulatory genes, THI2 (PH06) and THI3. We have also isolated a hybrid plasmid, pTTRl, containing a 6.2-kb DNA fragment from an S. cerevisiae genomic library which complements thiamine auxotrophy in the thi3 mutant. This gene was localized on a 3.0-kb ClaI-BglII fragment in the subclone pTTR5. Complementation of the activities for thiamine metabolism in the thi3 mutant transformed by some plasmids with the THI3 gene was also examined.

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“…Evidence to support this regulation includes the finding that ΔTHI2 and ΔTHI3 mutations significantly reduce the expression levels of thiamine biosynthesis "THI" genes (i.e., THI4, THI5, THI6, PHO3, and THI80) (10)(11)(12). Further studies revealed that Thi2p contains an N-terminal Zn 2 -Cys 6 domain that binds DNA upstream of the THI genes (13) and that Thi3p harbors a motif, Gly-Asp-Gly-X 24 -27 -Asn-Asn (where X is any amino acid residue), which is an apparent TPP sensor in the regulation of THI gene expression (14).…”

mentioning

confidence: 99%

ThiN as a Versatile Domain of Transcriptional Repressors and Catalytic Enzymes of Thiamine Biosynthesis

et al. 2017

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Thiamine biosynthesis is commonly regulated by a riboswitch mechanism; however, the enzymatic steps and regulation of this pathway in archaea are poorly understood. Haloferax volcanii, one of the representative archaea, uses a eukaryote-like Thi4 (thiamine thiazole synthase) for the production of the thiazole ring and condenses this ring with a pyrimidine moiety synthesized by an apparent bacterium-like ThiC (2-methyl-4-amino-5-hydroxymethylpyrimidine [HMP] phosphate synthase) branch. Here we found that archaeal Thi4 and ThiC were encoded by leaderless transcripts, ruling out a riboswitch mechanism. Instead, a novel ThiR transcription factor that harbored an N-terminal helix-turn-helix (HTH) DNA binding domain and C-terminal ThiN (TMP synthase) domain was identified. In the presence of thiamine, ThiR was found to repress the expression of thi4 and thiC by a DNA operator sequence that was conserved across archaeal phyla. Despite having a ThiN domain, ThiR was found to be catalytically inactive in compensating for the loss of ThiE (TMP synthase) function. In contrast, bifunctional ThiDN, in which the ThiN domain is fused to an N-terminal ThiD (HMP/HMP phosphate [HMP-P] kinase) domain, was found to be interchangeable for ThiE function and, thus, active in thiamine biosynthesis. A conserved Met residue of an extended ␣-helix near the active-site His of the ThiN domain was found to be important for ThiDN catalytic activity, whereas the corresponding Met residue was absent and the ␣-helix was shorter in ThiR homologs. Thus, we provide new insight into residues that distinguish catalytic from noncatalytic ThiN domains and reveal that thiamine biosynthesis in archaea is regulated by a transcriptional repressor, ThiR, and not by a riboswitch.IMPORTANCE Thiamine pyrophosphate (TPP) is a cofactor needed for the enzymatic activity of many cellular processes, including central metabolism. In archaea, thiamine biosynthesis is an apparent chimera of eukaryote-and bacterium-type pathways that is not well defined at the level of enzymatic steps or regulatory mechanisms. Here we find that ThiN is a versatile domain of transcriptional repressors and catalytic enzymes of thiamine biosynthesis in archaea. Our study provides new insight into residues that distinguish catalytic from noncatalytic ThiN domains and reveals that archaeal thiamine biosynthesis is regulated by a ThiN domain transcriptional repressor, ThiR, and not by a riboswitch.

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Cloning and characteristics of a positive regulatory gene, THI2 (PHO6), of thiamin biosynthesis in Saccharomyces cerevisiae

Cited by 49 publications

References 11 publications

Regulation of Gene Expression by Cofactors Derived from B Vitamins.

Regulation of Gene Expression by Cofactors Derived from B Vitamins.

A positive regulatory gene, THI3, is required for thiamine metabolism in Saccharomyces cerevisiae

ThiN as a Versatile Domain of Transcriptional Repressors and Catalytic Enzymes of Thiamine Biosynthesis

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