Effect of growth rate on the amounts of ribosomal and transfer ribonucleic acids in yeast

Waldron, Clive; Lacroute, François

doi:10.1128/jb.122.3.855-865.1975

Cited by 271 publications

(164 citation statements)

References 28 publications

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“…Because a yeast cell (cell volume 30-60 fl) contains 2-3 9 10 6 tRNA molecules (molecular weight ca. 25 kDa) (Waldron & Lacroute 1975;Jorgensen et al 2002), cellular concentration of tRNA is roughly estimated as 1-4 lg/lL. Therefore, I guess that the result led from this assay is not far apart from physiological condition.…”

Section: Discussionmentioning

confidence: 93%

Novel tRNA function in amino acid sensing of yeast Tor complex1

Kamada

2017

Genes to Cells

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TOR complex1 (TORC1), a master regulator of cell growth, is regulated by amino acids. Amino acids are fundamental nutrients, and 20 species of amino acids building proteins are not interchangeable with each other. Therefore, TORC1 should sense each amino acid individually. Mammalian mTORC1 is controlled by Rag GTPases and their regulators. However, Rag factors are dispensable for amino acid sensing by TORC1 in the budding yeast, suggesting an alternative mechanism of TORC1 regulation. Here, genetic investigation discovered the involvement of (aminoacyl-)tRNA ((aa-)tRNA) in TORC1 regulation. Biochemical TORC1 assay also showed that tRNA directly inhibits TORC1 kinase activity. Reducing cellular tRNA molecule desensitizes TORC1 inactivation by nitrogen starvation in vivo. Based on these results, I propose a model of the TORC1 regulatory mechanism in which free tRNA released from protein synthesis under amino acid starvation inhibits TORC1 activity. Therefore, TORC1 uses tRNA-mediated mechanism and Rag factors in parallel to sense intracellular amino acids.

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Section: Discussionmentioning

confidence: 93%

Novel tRNA function in amino acid sensing of yeast Tor complex1

Kamada

2017

Genes to Cells

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“…The fraction of the PPRI message at equilibrium would therefore be equal to 1.2 x 10-i. Taking into account the poly(A) RNA content of a yeast cell (0.016 pg, Waldron and Lacroute, 1975) and the size of the PPRI message (2.9 kb), it follows that there are 0.12 :1 0.1 PPRI mRNA molecule per cell.…”

Section: Discussionmentioning

confidence: 99%

“…Spheroplasts were prepared as described by Chevallier et al (1980) and lysed in acetate buffer (1 mM sodium acetate, 5 mM NaCl, 0.1 mM magnesium acetate, pH 5.1) containing 0.507o (w/v) SDS and 0.207o diethylpyrocarbonate. Total cellular RNA was purified from the lysed spheroplasts by three extractions with an equal volume of buffer-saturated phenol (Waldron and Lacroute, 1975). The poly(A) fractionation of RNA was carried out according to Fraser (1975).…”

Section: Cloning In Phage M13 Mp7mentioning

confidence: 99%

In vivo transcription of a eukaryotic regulatory gene.

Losson¹,

Fuchs²,

Lacroute³

1983

The EMBO Journal

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The PPR1 gene encodes the positive regulator of the URA1 and URA3 genes in yeast. Its transcription product is a 2.9‐kb polyadenylated RNA with an extremely short half‐life of 1 min. The induced or non‐induced cell contains approximately 0.1 molecules of PPR1 RNA, a constitutive level which is not altered by changing the number of structural genes to be regulated. The DNA sequence of a 399‐bp AccI‐Bg/II fragment including 180 nucleotides of the 5′‐flanking region of the gene PPR1 has been determined. By S1 mapping we present evidence that the 5′ non‐coding region of the PPR1 mRNA is heterogeneous in length. The 5′ termini of the different RNAs map 20, 36, 45 and 50 nucleotides upstream from the translation start codon. The sequence indicates that the only open translation phase begins with an AUG codon that is preceded by two out‐of‐frame AUG triplets. This particular structure of the 5′‐terminal sequence of the transcripts of PPR1 is discussed in relation to both their stability and translational efficiency.

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“…Finally, fluorescent in situ hybridization (FISH) of rRNA with fluorophore-labelled probes can be used to determine growth rate of individual biofilm cells by CLSM. In several microorganisms, the number of ribosomes is correlated with the growth rate in exponential phase (Kjeldgaard & Kurland, 1963;Waldron & Lacroute, 1975;Poulsen et al, 1993;Møller et al, 1995). A standard correlation between growth rate and ribosomal content as measured by quantitative FISH has been applied to the exponential and stationary phases of bacteria (Yang et al, 2008).…”

Section: Determination Of Growth Rate With Clsmmentioning

confidence: 99%

Saccharomyces cerevisiae— a model to uncover molecular mechanisms for yeast biofilm biology

Bojsen

Andersen

Regenberg

2012

FEMS Immunol Med Microbiol

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Microbial biofilms can be defined as multi-cellular aggregates adhering to a surface and embedded in an extracellular matrix (ECM). The nonpathogenic yeast, Saccharomyces cerevisiae, follows the common traits of microbial biofilms with cell-cell and cell-surface adhesion. S. cerevisiae is shown to produce an ECM and respond to quorum sensing, and multi-cellular aggregates have lowered susceptibility to antifungals. Adhesion is mediated by a family of cell surface proteins of which Flo11 has been shown to be essential for biofilm development. FLO11 expression is regulated via a number of regulatory pathways including the protein kinase A and a mitogen-activated protein kinase pathway. Advanced genetic tools and resources have been developed for S. cerevisiae including a deletion mutant-strain collection in a biofilm-forming strain background and GFP-fusion protein collections. Furthermore, S. cerevisiae biofilm is well applied for confocal laser scanning microscopy and fluorophore tagging of proteins, DNA and RNA. These techniques can be used to uncover the molecular mechanisms for biofilm development, drug resistance and for the study of molecular interactions, cell response to environmental cues, cell-to-cell variation and niches in S. cerevisiae biofilm. Being closely related to Candida species, S. cerevisiae is a model to investigate biofilms of pathogenic yeast.

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Effect of growth rate on the amounts of ribosomal and transfer ribonucleic acids in yeast

Cited by 271 publications

References 28 publications

Novel tRNA function in amino acid sensing of yeast Tor complex1

Novel tRNA function in amino acid sensing of yeast Tor complex1

In vivo transcription of a eukaryotic regulatory gene.

Saccharomyces cerevisiae— a model to uncover molecular mechanisms for yeast biofilm biology

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