The glycosylation of phosphoglucomutase is modulated by carbon source and heat shock in Saccharomyces cerevisiae.

Background: Tryptamine, a biogenic monoamine that is present in trace levels in the mammalian central nervous system, has probable roles as a neurotransmitter and/or a neuromodulator and may be associated with various neuropsychiatric disorders. One of the ways tryptamine may affect the body is by the competitive inhibition of the attachment of tryptophan to tryptophanyl tRNA synthetases. Methods: This study has explored the effects of tryptamine on growth of six yeast species (Saccharomyces cerevisiae, Candida glabrata, C. krusei, C. dubliniensis, C. tropicalis and C. lusitaniae) in media with glucose or ethanol as the carbon source, as well as recovery of growth inhibition by the addition of tryptophan. Results: Tryptamine was found to have an inhibitory effect on respiratory growth of all yeast species when grown with ethanol as the carbon source. Tryptamine also inhibited fermentative growth of Saccharomyces cerevisiae, C. krusei and C. tropicalis with glucose as the carbon source. In most cases the inhibitory effects were reduced by added tryptophan. Conclusion: The results obtained in this study are consistent with tryptamine competing with tryptophan to bind mitochondrial and cytoplasmic tryptophanyl tRNA synthetases in yeast: effects on mitochondrial and cytoplasmic protein synthesis can be studied as a function of growth with glucose or ethanol as a carbon source. Of the yeast species tested, there is variation in the sensitivity to tryptamine and the rescue by tryptophan. The current study suggests appropriate yeast strains and approaches for further studies.

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Is 20S RNA naked?

Molecular and Cellular Biology ◽

10.1128/mcb.11.5.2905 ◽

1991 ◽

Vol 11 (5) ◽

pp. 2905-2908 ◽

Cited By ~ 20

Author(s):

W R Widner ◽

Y Matsumoto ◽

R B Wickner

Keyword(s):

Saccharomyces Cerevisiae ◽

Life Cycle ◽

Heat Shock ◽

Heat Shock Protein ◽

Rna Virus ◽

Circular Rna ◽

Ribonucleoprotein Particle ◽

Multiple Copies

The 20S RNA of Saccharomyces cerevisiae is a single-stranded, circular RNA virus. A previous study suggested that this RNA is part of a 32S ribonucleoprotein particle, being associated with multiple copies of a 23-kilodalton protein. We show here that this protein is, in fact, the chromosome-encoded heat shock protein Hsp26. Furthermore, it is apparently not associated with 20S RNA and plays no obvious role in the life cycle of the virus.

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DNA damage and heat shock dually regulate genes in Saccharomyces cerevisiae.

Molecular and Cellular Biology ◽

10.1128/mcb.6.1.90 ◽

1986 ◽

Vol 6 (1) ◽

pp. 90-96 ◽

Cited By ~ 39

Author(s):

T McClanahan ◽

K McEntee

Keyword(s):

Saccharomyces Cerevisiae ◽

Dna Damage ◽

Heat Shock ◽

Shock Treatment ◽

Northern Hybridization ◽

Heat Shock Treatment ◽

Base Pairs ◽

S1 Nuclease ◽

Hsp70 Family ◽

Transcriptional Start Sites

Two Saccharomyces cerevisiae genes isolated in a differential hybridization screening for DNA damage regulation (DDR genes) were also transcriptionally regulated by heat shock treatment. A 0.45-kilobase transcript homologous to the DDRA2 gene and a 1.25-kilobase transcript homologous to the DDR48 gene accumulated after exposure of cells to 4-nitroquinoline-1-oxide (NQO; 1 to 1.5 microgram/ml) or brief heat shock (20 min at 37 degrees C). The DDRA2 transcript, which was undetectable in untreated cells, was induced to high levels by these treatments, and the DDR48 transcript increased more than 10-fold as demonstrated by Northern hybridization analysis. Two findings argue that dual regulation of stress-responsive genes is not common in S. cerevisiae. First, two members of the heat shock-inducible hsp70 family of S. cerevisiae, YG100 and YG102, were not induced by exposure to NQO. Second, at least one other DNA-damage-inducible gene, DIN1, was not regulated by heat shock treatment. We examined the structure of the induced RNA homologous to DDRA2 after heat shock and NQO treatments by S1 nuclease protection experiments. Our results demonstrated that the DDRA2 transcript initiates equally frequently at two sites separated by 5 base pairs. Both transcriptional start sites were utilized when cells were exposed to either NQO or heat shock treatment. These results indicate that DDRA2 and DDR48 are members of a unique dually regulated stress-responsive family of genes in S. cerevisiae.

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hsp26 of Saccharomyces cerevisiae is related to the superfamily of small heat shock proteins but is without a demonstrable function.

Molecular and Cellular Biology ◽

10.1128/mcb.9.11.5265 ◽

1989 ◽

Vol 9 (11) ◽

pp. 5265-5271 ◽

Cited By ~ 59

Author(s):

R E Susek ◽

S L Lindquist

Keyword(s):

Saccharomyces Cerevisiae ◽

Heat Shock ◽

Heat Shock Protein ◽

Mutational Analysis ◽

Small Heat Shock Protein ◽

Major Effect ◽

Selfish Dna ◽

Dna Elements ◽

Cloned Gene ◽

Shock Proteins

Analysis of the cloned gene confirms that hsp26 of Saccharomyces cerevisiae is a member of the small heat shock protein superfamily. Previous mutational analysis failed to demonstrate any function for the protein. Further experiments presented here demonstrate that hsp26 has no obvious regulatory role and no major effect on thermotolerance. It is possible that the small heat shock protein genes originated as primitive viral or selfish DNA elements.

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Identification of an upstream activating sequence and an upstream repressible sequence of the pyruvate kinase gene of the yeast Saccharomyces cerevisiae

Molecular and Cellular Biology ◽

10.1128/mcb.9.2.442-451.1989 ◽

1989 ◽

Vol 9 (2) ◽

pp. 442-451

Author(s):

M Nishizawa ◽

R Araki ◽

Y Teranishi

Keyword(s):

Saccharomyces Cerevisiae ◽

Carbon Source ◽

Pyruvate Kinase ◽

Transcriptional Activation ◽

Regulatory Elements ◽

Noncoding Region ◽

Yeast Cells ◽

Kinase Gene ◽

Yeast Saccharomyces Cerevisiae ◽

Upstream Activating Sequence

To clarify carbon source-dependent control of the glycolytic pathway in the yeast Saccharomyces cerevisiae, we have initiated a study of transcriptional regulation of the pyruvate kinase gene (PYK). By deletion analysis of the 5'-noncoding region of the PYK gene, we have identified an upstream activating sequence (UASPYK1) located between 634 and 653 nucleotides upstream of the initiating ATG codon. The promoter activity of the PYK 5'-noncoding region was abolished when the sequence containing the UASPYK1 was deleted from the region. Synthetic UASPYK1 (26mer), in either orientation, was able to restore the transcriptional activity of UAS-depleted mutants when placed upstream of the TATA sequence located at -199 (ATG as +1). While the UASPYK1 was required for basal to intermediate levels of transcriptional activation, a sequence between -714 and -811 was found to be necessary for full activation. On the other hand, a sequence between -344 and -468 was found to be responsible for transcriptional repression of the PYK gene when yeast cells were grown on nonfermentable carbon sources. This upstream repressible sequence also repressed transcription, although to a lesser extent, when glucose was present in the medium. The possible mechanism for carbon source-dependent regulation of PYK expression through these cis-acting regulatory elements is discussed.

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Repetitive Dictyostelium heat-shock promotor functions in Saccharomyces cerevisiae

Molecular and Cellular Biology ◽

10.1128/mcb.4.4.591-598.1984 ◽

1984 ◽

Vol 4 (4) ◽

pp. 591-598

Author(s):

J Cappello ◽

C Zuker ◽

H F Lodish

Keyword(s):

Saccharomyces Cerevisiae ◽

Heat Shock ◽

Nucleotide Sequence ◽

Dna Sequence ◽

Base Pair ◽

Shock Treatment ◽

Yeast Cells ◽

Genomic Segment ◽

Heat Shock Treatment ◽

Shock Response

The Dictyostelium genome contains 40 copies of a 4.7-kilobase repetitive and apparently transposable DNA sequence (DIRS-1) and about 250 smaller elements that appear to be deletions or rearrangements of DIRS-1. Transcripts of these sequences are induced during differentiation and also by heat shock treatment of growing cells. We showed that one such cloned element, pB41.6 (2.5 kilobases) contains a nucleotide sequence identical to the Drosophila consensus heat shock promotor. To test whether this sequence might indeed control the expression of DIRS-1-related RNAs, we have cloned this genomic segment into yeast cells. In yeast cells, 41.6 directs synthesis of a 1.7-kilobase RNA that is induced at least 10-fold by heat shock. Transcription initiates at about 124 bases 3' of the putative promotor sequence and terminates within the 41.6 insert. A 381-base-pair subclone that contains the putative promotor sequence is sufficient to induce the heat shock response of 41.6 in yeast cells.

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Saccharomyces cerevisiae contains two functional citrate synthase genes

Molecular and Cellular Biology ◽

10.1128/mcb.6.6.1936-1942.1986 ◽

1986 ◽

Vol 6 (6) ◽

pp. 1936-1942

Author(s):

K S Kim ◽

M S Rosenkrantz ◽

L Guarente

Keyword(s):

Saccharomyces Cerevisiae ◽

Carbon Source ◽

Citrate Synthase ◽

Tricarboxylic Acid Cycle ◽

Nuclear Gene ◽

Tricarboxylic Acid ◽

Yeast Genome ◽

Gene Encoding ◽

Acid Cycle ◽

Nonfermentable Carbon Source

The tricarboxylic acid cycle occurs within the mitochondria of the yeast Saccharomyces cerevisiae. A nuclear gene encoding the tricarboxylic acid cycle enzyme citrate synthase has previously been isolated (M. Suissa, K. Suda, and G. Schatz, EMBO J. 3:1773-1781, 1984) and is referred to here as CIT1. We report here the isolation, by an immunological method, of a second nuclear gene encoding citrate synthase (CIT2). Disruption of both genes in the yeast genome was necessary to produce classical citrate synthase-deficient phenotypes: glutamate auxotrophy and poor growth on rich medium containing lactate, a nonfermentable carbon source. Therefore, the citrate synthase produced from either gene was sufficient for these metabolic roles. Transcription of both genes was maximally repressed in medium containing both glucose and glutamate. However, transcription of CIT1 but not of CIT2 was derepressed in medium containing a nonfermentable carbon source. The significance of the presence of two genes encoding citrate synthase in S. cerevisiae is discussed.

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Heat shock response of Saccharomyces cerevisiae mutants altered in cyclic AMP-dependent protein phosphorylation

Molecular and Cellular Biology ◽

10.1128/mcb.7.1.244-250.1987 ◽

1987 ◽

Vol 7 (1) ◽

pp. 244-250

Author(s):

D Y Shin ◽

K Matsumoto ◽

H Iida ◽

I Uno ◽

T Ishikawa

Keyword(s):

Heat Treatment ◽

Saccharomyces Cerevisiae ◽

Heat Shock ◽

Cyclic Amp ◽

Heat Shock Proteins ◽

Temperature Shift ◽

G1 Phase ◽

Shock Response ◽

Dependent Protein ◽

Shock Proteins

When Saccharomyces cerevisiae cells grown at 23 degrees C were transferred to 36 degrees C, they initiated synthesis of heat shock proteins, acquired thermotolerance to a lethal heat treatment given after the temperature shift, and arrested their growth transiently at the G1 phase of the cell division cycle. The bcy1 mutant which resulted in production of cyclic AMP (cAMP)-independent protein kinase did not synthesize the three heat shock proteins hsp72A, hsp72B, and hsp41 after the temperature shift. The bcy1 cells failed to acquire thermotolerance to the lethal heat treatment and were not arrested at the G1 phase after the temperature shift. In contrast, the cyr1-2 mutant, which produced a low level of cAMP, constitutively produced three heat shock proteins and four other proteins without the temperature shift and was resistant to the lethal heat treatment. The results suggest that a decrease in the level of cAMP-dependent protein phosphorylation results in the heat shock response, including elevated synthesis of three heat shock proteins, acquisition of thermotolerance, and transient arrest of the cell cycle.

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Heat shock factor can activate transcription while bound to nucleosomal DNA in Saccharomyces cerevisiae

Molecular and Cellular Biology ◽

10.1128/mcb.14.1.189-199.1994 ◽

1994 ◽

Vol 14 (1) ◽

pp. 189-199

Author(s):

D S Pederson ◽

T Fidrych

Keyword(s):

Saccharomyces Cerevisiae ◽

Heat Shock ◽

Transcription Initiation ◽

Heat Shock Factor ◽

Micrococcal Nuclease ◽

Nucleosome Assembly ◽

Heat Shock Element ◽

Yeast Saccharomyces Cerevisiae ◽

Nucleosomal Dna

After each round of replication, new transcription initiation complexes must assemble on promoter DNA. This process may compete with packaging of the same promoter sequences into nucleosomes. To elucidate interactions between regulatory transcription factors and nucleosomes on newly replicated DNA, we asked whether heat shock factor (HSF) could be made to bind to nucleosomal DNA in vivo. A heat shock element (HSE) was embedded at either of two different sites within a DNA segment that directs the formation of a stable, positioned nucleosome. The resulting DNA segments were coupled to a reporter gene and transfected into the yeast Saccharomyces cerevisiae. Transcription from these two plasmid constructions after induction by heat shock was similar in amount to that from a control plasmid in which HSF binds to nucleosome-free DNA. High-resolution genomic footprint mapping of DNase I and micrococcal nuclease cleavage sites indicated that the HSE in these two plasmids was, nevertheless, packaged in a nucleosome. The inclusion of HSE sequences within (but relatively close to the edge of) the nucleosome did not alter the position of the nucleosome which formed with the parental DNA fragment. Genomic footprint analyses also suggested that the HSE-containing nucleosome was unchanged by the induction of transcription. Quantitative comparisons with control plasmids ruled out the possibility that HSF was bound only to a small fraction of molecules that might have escaped nucleosome assembly. Analysis of the helical orientation of HSE DNA in the nucleosome indicated that HSF contacted DNA residues that faced outward from the histone octamer. We discuss the significance of these results with regard to the role of nucleosomes in inhibiting transcription and the normal occurrence of nucleosome-free regions in promoters.

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Identification of cis and trans components of a novel heat shock stress regulatory pathway in Saccharomyces cerevisiae

Molecular and Cellular Biology ◽

10.1128/mcb.13.1.248-256.1993 ◽

1993 ◽

Vol 13 (1) ◽

pp. 248-256

Author(s):

N Kobayashi ◽

K McEntee

Keyword(s):

Saccharomyces Cerevisiae ◽

Stress Response ◽

Heat Shock ◽

Reporter Gene ◽

Heat Shock Element ◽

Cis Acting ◽

Heat Shock Stress ◽

Specific Complex ◽

Cis And Trans ◽

Shock Stress

The stress-responsive DDR2 gene (previously called DDRA2) of Saccharomyces cerevisiae is transcribed at elevated levels following stress caused by heat shock or DNA damage. Previously, we identified a 51-bp promoter fragment, oligo31/32, which conferred heat shock inducibility on the heterologous CYC1-lacZ reporter gene in S. cerevisiae (N. Kobayashi and K. McEntee, Proc. Natl. Acad. Sci. USA 87:6550-6554, 1990). Using a series of synthetic oligonucleotides, we have identified a pentanucleotide, CCCCT (C4T), as an essential component of this stress response sequence. This element is not a binding site for the well-characterized heat shock transcription factor which recognizes a distinct cis-acting heat shock element in the promoters of many heat shock genes. Here we demonstrate the ability of oligonucleotides containing the C4T sequence to confer heat shock inducibility on the reporter gene and show that the presence of two such elements produces more than additive effects on induction. Gel retardation experiments have been used to demonstrate specific complex formation between C4T-containing fragments and one or more yeast proteins. Formation of these complexes was not competed by fragments containing mutations in the C4T sequence nor by heat shock element-containing competitor DNAs. Fragments containing the C4T element bound to a single 140-kDa polypeptide, distinct from heat shock transcription factors in yeast crude extracts. These experiments identify key cis- and trans-acting components of a novel heat shock stress response pathway in S. cerevisiae.

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