Two new genes controlling the constitutive acid phosphatase synthesis in Saccharomyces cerevisiae

Toh‐e, Akio; Kakimoto, Sei-ichiro; Oshima, Yasuji

doi:10.1007/bf00332380

Cited by 18 publications

(13 citation statements)

References 4 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…Transformants of the f/ti2(,dod) mutant, K272-7c, harboring the low-copy numbered vector plasmid YCp50, pRS316 and four plasmid (pTSR1, pTSR2, pTSR3 and pTSR4) were cultured in the absence of thiamin and assayed for T-rAPase activity ( Table I). The T-rAPasc activity in these transformants harboring the T1//2(PH06) gene was less active than the wild-type strain and its activity in the recipient cells transformed with the control plasmid, YCp50 and pRS3 16, were as markedly low as that of the dti2(phu6) mutant, OS&MS [2]. Furthermore, the activity in these transformants with pTSR1, pTSR2 and pTSR3 was repressed by thiamin (_ 3 x lo-' M) in the minimal medium in the same manner as that of the wild-type strain.…”

Section: Expwssion Of Ffte Uctivifies Of T-rapuse Und Cii-mentioning

confidence: 87%

“…and after centrifugation at 28.000 x g for 30 min 111~ supernatant was used as a crude extract. Overall thiamin-synthesizing enzyme activity from hydroxymcthylpyrimidine and hydroxycthylthiazolc and the activity of thiamin phosphate pyrophosphorylasc were assayed as previously described [3] The thi2(pho6) mutant defective in a regulatory gene for the synthesis of T-rAPase [2] was auxotrophic for thiamin, and the activities of enzymes involved in thiamin synthesis were markedly low in the crude extract from the thi2(pho6) mutant [3]. However, as shown in Fig.…”

Section: Sfraim Mtd Rrrdiumentioning

confidence: 99%

“…The thiamin-repressible acid phosphatase (T-rAPase) in the periplusmic space of Smdtarottt_wes cerew3iue is coded by the PH03 gene [ 1] and requires two additional complementary genes, PHU6 and PH07, for its synthesis [2]. In a previous paper [3], we indicated that a p/to6 mutant was auxotrophic for thiamin and the PH06 gene was involved in the regula'tion of the synthesis not only of T-rAPase but also of enzymes synthesizing thiamin monophosphate from 2-methyl-4-amino&hy-droxymethyl-pyrimidine (hydroxymethylpyrimidine) and 4-methyl-5-fl-hydroxyethylthiazole (hydroxyethylthiazole) in S. cerevisiae [4-71. Our previous study also showed that the activities of these enzymes are coordinately repressed by exogenous thiamin and that these activities were decreased in correlation with the insrease in the intracellular thiamin pyrophosphate level in yeast cells [3].…”

Section: Introductionmentioning

confidence: 99%

See 2 more Smart Citations

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

et al. 1992

View full text Add to dashboard Cite

A rlri@lrob) mutant or Succhurorr~_w,~ ccwi.siue. dclixtivc in the expression or structural genes !'or ihiamin-repressible acid phosphatasc and enzymes involved in thiumin biosynthesis. was found to relain sufiicient thiamin transport activhy. The transport activity was repressed by thiamin in growth mrdium. WC isolated rrom a S. rcrcvisbe gcnomic library two hybrid plasmids. pTSR I and pTSRZ, containing 10.2-and ILO-kilobase (kb) DNA frirgmenls. respectively, \vhich complcmcni the 1lri-'oll,n6) mutaiion of S. wrdsiue. This pnc was localized on a G.O-kb C'fal-C/u! lkagmcnt in the subclonc pTSR3. Complementation of the enzyme activilies for lhiamin metabolism in the rlti2@lru6) mutant transformed by some plasmids wit!1 the THf?(PHC?6) gene was also examined.

show abstract

Section: Expwssion Of Ffte Uctivifies Of T-rapuse Und Cii-mentioning

confidence: 87%

Section: Sfraim Mtd Rrrdiumentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

See 1 more Smart Citation

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

et al. 1992

View full text Add to dashboard Cite

show abstract

“…On the other hand, apho6 mutant defective in the regulatory gene for the synthesis of periplasmic T-rAPase encoded by the PHO3 gene (26,27) is auxotrophic for thiamine, and the enzyme activities involved in thiamine synthesis described above are markedly low in crude extracts of this mutant (9). This indicated that the expression of structural genes for T-rAPase and the enzymes involved in thiamine biosynthesis was regulated positively by PHO6.…”

mentioning

confidence: 99%

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

et al. 1992

View full text Add to dashboard Cite

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.

show abstract

“…PHO2 mutants lack only the repressible APase (26,28). PHO6 and PHO7 affect expression of PHO3, the "constitutive" APase (23), and PHO9 encodes a protease involved in AlkPase maturation (9).…”

mentioning

confidence: 99%

Regulation of repressible acid phosphatase gene transcription in Saccharomyces cerevisiae.

Lemire¹,

Willcocks²,

Halvorson³

et al. 1985

Mol. Cell. Biol.

View full text Add to dashboard Cite

We examined the genetic system responsible for transcriptional regulation of repressible acid phosphatase (APase; orthophosphoric-monoester phosphohydrolase [acid optimum, EC 3. (7,21,29); and two enzymes, alkaline phosphatase (AlkPase) and polyphosphatase, located in the yeast vacuole, which hydrolyze polyphosphate (7, 9). These enzymes regulate intracellular concentrations of Pi and maintain cellular homeostasis (7, 8) by a cyclic pathway of polyphosphate synthesis and degradation (7). They are themselves regulated by external growth concentrations of Pi (6,7,9,10).This family of enzymes is a dispersed system composed of numerous structural and regulatory genes (Table 1). There are four known structural genes encoding APase (PHO3, PHO5, PHOJO, PHOII), genes for AlkPase (PHO8) and Pi transport (PHO84), and eight genes affecting their expression (9, 23-28). Recessive mutations in PHO4 and PHO81 block the derepression of repressible APase and AlkPase, in addition to Pi uptake (26, 28), whereas recessive mutations in PHO80 and PHO85 result in their constitutive expression. PHO2 mutants lack only the repressible APase (26, 28). PHO6 and PHO7 affect expression of PHO3, the "constitutive" APase (23), and PHO9 encodes a protease involved in AlkPase maturation (9).On the basis of genetic analysis of various double mutants and the discovery of a cis-dominant constitutive mutant (PHO82) contiguous to PH04 (26), Toh-e et al. originally proposed a role for the phosphatase regulatory genes involving transcriptional regulation via the sequential functioning of their products (28), reminiscent of control of phage * Corresponding author.development by repressor and antirepressor proteins. PHO4 was considered to be a positive regulator essential for transcription of PHOS and PHO8. The PHO80 and PHO85 proteins form a repressor under negative control of PHO81.In the presence of Pi, this repressor blocks transcription of PHO4 by binding to an adjacent site pho82. In the absence of P1 it dissociates, allowing for synthesis of PHO4 which, in turn, associates with sites adjacent to PHOS and PHO8 and activates transcription. For APase, this occurs in concert with PHO2 at the pho83 locus adjacent to PHOS.More recent genetic studies, however, are inconsistent with this model. First, fine-structure meiotic mapping situates the pho82 site in a narrow region within the PHO4 locus (22). The existence of temperature-sensitive and nonsense suppressible mutations in PHO4 argues strongly that this locus encodes a protein (22). The fact that two PHO4 sites flank pho82 therefore argues against a model defining the pho82 site as an operator of PHO4. Moreover, APase activity shown by PHO82 PHO4Ipho82 (wild-type) pho4 diploids grown under repressed conditions varied depending on the combination of PHO82 and pho4 alleles, unlike the PHO82 homozygous diploids (22).These findings led Toh-e et al. to propose a new model wherein the regulatory factors function simultaneously by direct molecular interaction (22). In its simplest form, this new model stat...

show abstract

Two new genes controlling the constitutive acid phosphatase synthesis in Saccharomyces cerevisiae

Abstract: Two new classes of mutants, phoF and phoG, lacking the constitutive acid phosphatase activity, were isolated. They both complemented each other and the phoC mutation. No linkage was detected among these three complementary genes.

Cited by 18 publications

References 4 publications

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

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

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

Regulation of repressible acid phosphatase gene transcription in Saccharomyces cerevisiae.

Contact Info

Product

Resources

About