RNA and homology mapping of two DNA fragments with repressible acid phosphatase genes from Saccharomyces cerevisiae.

Andersen, N; Thill, G P; Kramer, Richard A.

doi:10.1128/mcb.3.4.562

Cited by 26 publications

(14 citation statements)

References 23 publications

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“…Recently, another cross-reactive APase polypeptide of 57,000 daltons was identified and shown to be the product of a second gene present on the 8-kb EcoRl S. cerevisiae fragment (1,30). Presumably, the region of weak homology on the 8-kb EcoRI fragment to the p56 and p60 genes observed by heteroduplex analysis (1) denotes the structural gene-coding region of the p57 gene.…”

Section: Discussionmentioning

confidence: 99%

“…We examined this possibility by characterizing the Nterminus of in vitro synthesized p60 polypeptide, which should retain an intact leader sequence because of the low processing activity endogenous to our cell-free translation system. The p60 polypeptide was 1 (Fig. 6C).…”

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confidence: 99%

“…The structural genes that encode the p60 and p56 proteins have recently been cloned on EcoRI restriction fragments of 8 and 5 kilobases (kb), respectively (20), and characterized by in vitro and in vivo analyses (30). The 8-kb EcoRI restriction fragment was also found to contain another APase gene, (p57), which is probably the genetically identified "constitutive" PH03 gene (1,30).…”

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Comparative analysis of the 5'-end regions of two repressible acid phosphatase genes in Saccharomyces cerevisiae.

Thill¹,

Kramer²,

Turner³

et al. 1983

Mol. Cell. Biol.

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The nucleotide sequence of 5'-noncoding and N-terminal coding regions of two coordinately regulated, repressible acid phosphatase genes from Saccharomyces cerevisiae were determined. These unlinked genes encode different, but structurally related polypeptides of molecular weights 60,000 and 56,000. The DNA sequences of their 5'-flanking regions show stretches of extensive homology upstream of, and surrounding, a "TATA" sequence and in a region in which heterogeneous 5' ends of the p60 mRNA were mapped. The predicted amino acid sequences encoded by the N-terminal regions of both genes were confirmed by determination of the amino acid sequence of the native exocellular acid phosphatase and the partial sequence of the presecretory polypeptide sythesized in a cellfree protein synthesizing system. The N-terminal region of the p60 polypeptide was shown to be characterized by a hydrophobic 17-amino acid signal polypeptide which is absent in the native exocellular protein and thought to be necessary for acid phosphatase secretion.

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Section: Discussionmentioning

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Comparative analysis of the 5'-end regions of two repressible acid phosphatase genes in Saccharomyces cerevisiae.

Thill¹,

Kramer²,

Turner³

et al. 1983

Mol. Cell. Biol.

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“…In contrast, APase mRNAs accumulated in its absence or presence. The control culture showed a typical biphasic appearance of mRNA, oscillating over hour 1 protein synthesis is not required for activating APase transcription but is necessary for the feedback and autoregulatory repression responsible for early mRNA oscillations.…”

Section: Methodsmentioning

confidence: 99%

Regulation of repressible acid phosphatase gene transcription in Saccharomyces cerevisiae.

Lemire¹,

Willcocks²,

Halvorson³

et al. 1985

Mol. Cell. Biol.

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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...

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“…This gene family is composed of at least three structural genes (PHO3, PHO5, PHO11), all coding for secreted acid phosphatases (13)(14)(15)(16)(17). The interplay of several regulatory proteins leads to differential expression of these genes with respect to the concentration of inorganic phosphate (Pi).…”

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The yeast PHO5 promoter: phosphate-control elements and sequences mediating mRNA start-site selection.

Rudolph¹,

Hinnen²

1987

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

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Transcription of PHOS is strongly regulated in response to the level of inorganic phosphate (PI) present in the growth medium. We have identified elements required for PHOS expression by analyzing small deletions in the PH05 promoter on chromosome II. The results reveal three functionally different components of the PH05 promoter: (i) regulatory regions, (ii) a "TATA" element, and (iii) specific mRNA initiation sites. The regulatory regions contain related 19-base-pair (bp) dyad sequences acting as phosphate-controlled upstream activation sites (UASps). These UASps mediate the transcriptional activation of PH05 observed in low Pi conditions. The unlinked but coordinately regulated PHOII promoter contains a single copy of an almost identical dyad sequence, suggesting that there is a common regulatory UASp for both genes. A TATA element is absolutely required for detectable PH05 transcription. Specific purine-pyrimidine motifs (RRYRR) (R = purine and Y = pyrimidine) serve as PH05 mRNA initiation sites, but only if they lie 55-110 bp downstream of a functional TATA element. Such an "initiation window" is not found in higher eukaryotes and implies mechanistic differences in the transcription machineries between yeast and higher eukaryotes.Faithful in vivo transcription of most eukaryotic genes by RNA polymerase II requires at least two distinct promoter elements. In higher eukaryotes, the first element is a highly conserved TATA motif usually found at a fixed distance of =30 base pairs (bp) upstream from the transcription initiation site (1). The second essential promoter element typically lies >100 bp upstream of the cap site and includes enhancers and gene-specific modulators (2-5).Transcription in yeast Saccharomyces cerevisiae depends on the same essential elements, but the TATA sequence is not in a strict positional relationship to the initiation site(s). Upstream elements can promote constitutive expression (6, 7) or act as regulatory sites, called upstream activation sites (UASs) (8), which interact with specific regulatory proteins.A well-studied example of an UAS is the region between the divergently transcribed GAL] and GALIO genes, UASG, to which a regulatory protein (GAL4) binds at four dyadsymmetric sequences mediating the galactose induction of both genes (9-11). Genetic analysis of the regulatory circuits involved in the GAL system reveals a striking similarity to those controlling the expression of the multigene family of acid phosphatases (12).This gene family is composed of at least three structural genes (PHO3, PHO5, PHO11), all coding for secreted acid phosphatases (13-17). The interplay of several regulatory proteins leads to differential expression of these genes with respect to the concentration of inorganic phosphate (Pi). The PH05 and PHOJJ genes are inducible by low Pi, whereas PH03 remains essentially unaffected or even decreases slightly under the same conditions (15). Coordinate transcriptional activation of PH05 and PHOJJ is thought to be executed by two positive factors, PH04 a...

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RNA and homology mapping of two DNA fragments with repressible acid phosphatase genes from Saccharomyces cerevisiae.

Cited by 26 publications

References 23 publications

Comparative analysis of the 5'-end regions of two repressible acid phosphatase genes in Saccharomyces cerevisiae.

Comparative analysis of the 5'-end regions of two repressible acid phosphatase genes in Saccharomyces cerevisiae.

Regulation of repressible acid phosphatase gene transcription in Saccharomyces cerevisiae.

The yeast PHO5 promoter: phosphate-control elements and sequences mediating mRNA start-site selection.

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