Partial characterization of an RNA component that copurifies with Saccharomyces cerevisiae RNase P

Saccharomyces cerevisiae cellular RNase P is composed of both protein and RNA components that are essential for activity. The isolated holoenzyme contains a highly structured RNA of 369 nucleotides that has extensive sequence similarities to the 286-nucleotide RNA associated with Schizosaccharomyces pombe RNase P but bears little resemblance to the analogous RNA sequences in procaryotes or S. cerevisiae mitochondria. Even so, the predicted secondary structure of S. cerevisiae RNA is strikingly similar to the bacterial phylogenetic consensus rather than to previously predicted structures of other eucaryotic RNase P RNAs.

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Characterization of the gene for fructose-1,6-bisphosphatase from Saccharomyces cerevisiae and Schizosaccharomyces pombe. Sequence, protein homology, and expression during growth on glucose.

Journal of Biological Chemistry ◽

10.1016/s0021-9258(18)68747-2 ◽

1988 ◽

Vol 263 (13) ◽

pp. 6051-6057 ◽

Cited By ~ 2

Author(s):

D T Rogers ◽

E Hiller ◽

L Mitsock ◽

E Orr

Keyword(s):

Saccharomyces Cerevisiae ◽

Schizosaccharomyces Pombe ◽

Protein Homology ◽

Fructose 1,6 Bisphosphatase

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Heterologous expression and characterization of Schizosaccharomyces pombe vacuolar carboxypeptidase Y in Saccharomyces cerevisiae

Current Genetics ◽

10.1007/s00294-002-0357-0 ◽

2002 ◽

Vol 42 (5) ◽

pp. 252-259 ◽

Cited By ~ 6

Author(s):

Kaoru Takegawa ◽

Sanae Tokudomi ◽

M. Shah Alam Bhuiyan ◽

Mitsuaki Tabuchi ◽

Yasuko Fujita ◽

...

Keyword(s):

Saccharomyces Cerevisiae ◽

Heterologous Expression ◽

Schizosaccharomyces Pombe ◽

Carboxypeptidase Y

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U2 small nuclear RNA is remarkably conserved between Schizosaccharomyces pombe and mammals

Molecular and Cellular Biology ◽

10.1128/mcb.8.12.5575-5580.1988 ◽

1988 ◽

Vol 8 (12) ◽

pp. 5575-5580

Author(s):

P Brennwald ◽

G Porter ◽

J A Wise

Keyword(s):

Saccharomyces Cerevisiae ◽

Secondary Structure ◽

Schizosaccharomyces Pombe ◽

Small Nuclear Rna ◽

Primary Sequence ◽

Cloning And Sequencing ◽

Sequence Identity ◽

Human Counterpart ◽

Nuclear Rna ◽

Rna Blot Analysis

We report the molecular cloning and sequencing of the most abundant trimethylguanosine-capped small nuclear RNA from the fission yeast Schizosaccharomyces pombe, a highly conserved homolog of mammalian U2 small nuclear RNA. This RNA is 186 nucleotides in length, just 2 nucleotides shorter than its human counterpart; this is in contrast to Saccharomyces cerevisiae U2, which is 1,175 nucleotides long. Moreover, the secondary structure of Schizosaccharomyces pombe U2 is virtually identical to that of mammalian U2, including the 3' half of the RNA, which shows limited primary sequence identity. Northern (RNA) blot analysis revealed that the size of this RNA is conserved not only in fission yeasts but in many organisms, including other ascomycetes.

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Characterization of RPR1, an essential gene encoding the RNA component of Saccharomyces cerevisiae nuclear RNase P.

Molecular and Cellular Biology ◽

10.1128/mcb.11.2.721 ◽

1991 ◽

Vol 11 (2) ◽

pp. 721-730 ◽

Cited By ~ 50

Author(s):

J Y Lee ◽

C E Rohlman ◽

L A Molony ◽

D R Engelke

Keyword(s):

Saccharomyces Cerevisiae ◽

Essential Gene ◽

Rnase P ◽

Single Copy ◽

Temperature Sensitive ◽

Coding Region ◽

Gene Encoding ◽

Processing Steps ◽

Potential Precursor

RNA components have been identified in preparations of RNase P from a number of eucaryotic sources, but final proof that these RNAs are true RNase P subunits has been elusive because the eucaryotic RNAs, unlike the procaryotic RNase P ribozymes, have not been shown to have catalytic activity in the absence of protein. We previously identified such an RNA component in Saccharomyces cerevisiae nuclear RNase P preparations and have now characterized the corresponding, chromosomal gene, called RPR1 (RNase P ribonucleoprotein 1). Gene disruption experiments showed RPR1 to be single copy and essential. Characterization of the gene region located RPR1 600 bp downstream of the URA3 coding region on chromosome V. We have sequenced 400 bp upstream and 550 bp downstream of the region encoding the major 369-nucleotide RPR1 RNA. The presence of less abundant, potential precursor RNAs with an extra 84 nucleotides of 5' leader and up to 30 nucleotides of 3' trailing sequences suggests that the primary RPR1 transcript is subjected to multiple processing steps to obtain the 369-nucleotide form. Complementation of RPR1-disrupted haploids with one variant of RPR1 gave a slow-growth and temperature-sensitive phenotype. This strain accumulates tRNA precursors that lack the 5' end maturation performed by RNase P, providing direct evidence that RPR1 RNA is an essential component of this enzyme.

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Characterization of Schizosaccharomyces pombe Malate Permease by Expression in Saccharomyces cerevisiae

Applied and Environmental Microbiology ◽

10.1128/aem.67.9.4144-4151.2001 ◽

2001 ◽

Vol 67 (9) ◽

pp. 4144-4151 ◽

Cited By ~ 46

Author(s):

Carole Camarasa ◽

Frédérique Bidard ◽

Muriel Bony ◽

Pierre Barre ◽

Sylvie Dequin

Keyword(s):

Saccharomyces Cerevisiae ◽

Schizosaccharomyces Pombe ◽

Malic Acid ◽

Dicarboxylic Acids ◽

Acid Uptake ◽

Acid Transport ◽

Gene Encoding ◽

Simple Diffusion ◽

External Ph

ABSTRACT In Saccharomyces cerevisiae, l-malic acid transport is not carrier mediated and is limited to slow, simple diffusion of the undissociated acid. Expression in S. cerevisiae of the MAE1 gene, encodingSchizosaccharomyces pombe malate permease, markedly increased l-malic acid uptake in this yeast. In this strain, at pH 3.5 (encountered in industrial processes),l-malic acid uptake involves Mae1p-mediated transport of the monoanionic form of the acid (apparent kinetic parameters:V max = 8.7 nmol/mg/min;Km = 1.6 mM) and some simple diffusion of the undissociated l-malic acid (Kd = 0.057 min−1). As total l-malic acid transport involved only low levels of diffusion, the Mae1p permease was further characterized in the recombinant strain. l-Malic acid transport was reversible and accumulative and depended on both the transmembrane gradient of the monoanionic acid form and the ΔpH component of the proton motive force. Dicarboxylic acids with stearic occupation closely related to l-malic acid, such as maleic, oxaloacetic, malonic, succinic and fumaric acids, inhibitedl-malic acid uptake, suggesting that these compounds use the same carrier. We found that increasing external pH directly inhibited malate uptake, resulting in a lower initial rate of uptake and a lower level of substrate accumulation. In S. pombe, proton movements, as shown by internal acidification, accompanied malate uptake, consistent with the proton/dicarboxylate mechanism previously proposed. Surprisingly, no proton fluxes were observed during Mae1p-mediated l-malic acid import inS. cerevisiae, and intracellular pH remained constant. This suggests that, in S. cerevisiae, either there is a proton counterflow or the Mae1p permease functions differently from a proton/dicarboxylate symport.

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Functional Expression and Characterization of Schizosaccharomyces pombe Avt3p as a Vacuolar Amino Acid Exporter in Saccharomyces cerevisiae

PLoS ONE ◽

10.1371/journal.pone.0130542 ◽

2015 ◽

Vol 10 (6) ◽

pp. e0130542 ◽

Cited By ~ 8

Author(s):

Soracom Chardwiriyapreecha ◽

Kunio Manabe ◽

Tomoko Iwaki ◽

Miyuki Kawano-Kawada ◽

Takayuki Sekito ◽

...

Keyword(s):

Saccharomyces Cerevisiae ◽

Amino Acid ◽

Schizosaccharomyces Pombe ◽

Functional Expression

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Identification of the Aldo-Keto Reductase Responsible for d-galacturonic Acid Conversion to l-galactonate in Saccharomyces cerevisiae

Journal of Fungi ◽

10.3390/jof7110914 ◽

2021 ◽

Vol 7 (11) ◽

pp. 914

Author(s):

Dorthe Rippert ◽

Federica Linguardo ◽

Andreea Perpelea ◽

Mathias Klein ◽

Elke Nevoigt

Keyword(s):

Saccharomyces Cerevisiae ◽

Galacturonic Acid ◽

Homology Search ◽

Cell Factory ◽

Citrus Peel ◽

Beet Pulp ◽

By Products ◽

Sequence Similarities

d-galacturonic acid (d-GalUA) is the main constituent of pectin, a complex polysaccharide abundant in several agro-industrial by-products such as sugar beet pulp or citrus peel. During several attempts to valorise d-GalUA by engineering the popular cell factory Saccharomyces cerevisiae, it became obvious that d-GalUA is, to a certain degree, converted to l-galactonate (l-GalA) by an endogenous enzymatic activity. The goal of the current work was to clarify the identity of the responsible enzyme(s). A protein homology search identified three NADPH-dependent unspecific aldo-keto reductases in baker’s yeast (encoded by GCY1, YPR1 and GRE3) that show sequence similarities to known d-GalUA reductases from filamentous fungi. Characterization of the respective deletion mutants and an in vitro enzyme assay with a Gcy1 overproducing strain verified that Gcy1 is mainly responsible for the detectable reduction of d-GalUA to l-GalA.

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