Characterization of a gene encoding Trametes versicolor laccase A and improved heterologous expression in Saccharomyces cerevisiae by decreased cultivation temperature

1999 ◽  
Vol 52 (3) ◽  
pp. 393-400 ◽  
Author(s):  
P. Cassland ◽  
L. J. Jönsson
Keyword(s):  
Saccharomyces Cerevisiae ◽  
Heterologous Expression ◽  
Trametes Versicolor ◽  
Cultivation Temperature ◽  
Gene Encoding

scholarly journals Heterologous expression and characterization of bacterial 2-C-methyl-d-erythritol-4-phosphate pathway in Saccharomyces cerevisiae

2013 ◽  
Vol 97 (13) ◽  
pp. 5753-5769 ◽  
Author(s):  
Simon Carlsen ◽  
Parayil Kumaran Ajikumar ◽  
Luca Riccardo Formenti ◽  
Kang Zhou ◽  
Too Heng Phon ◽  
...  
Keyword(s):  
Saccharomyces Cerevisiae ◽  
Heterologous Expression

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

2002 ◽  
Vol 42 (5) ◽  
pp. 252-259 ◽  
Author(s):  
Kaoru Takegawa ◽  
Sanae Tokudomi ◽  
M. Shah Alam Bhuiyan ◽  
Mitsuaki Tabuchi ◽  
Yasuko Fujita ◽  
...  
Keyword(s):  
Saccharomyces Cerevisiae ◽  
Heterologous Expression ◽  
Schizosaccharomyces Pombe ◽  
Carboxypeptidase Y

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

1991 ◽  
Vol 11 (2) ◽  
pp. 721-730 ◽  
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.

scholarly journals Characterization of Schizosaccharomyces pombe Malate Permease by Expression in Saccharomyces cerevisiae

2001 ◽  
Vol 67 (9) ◽  
pp. 4144-4151 ◽  
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.

scholarly journals Cloning and characterization of DST2, the gene for DNA strand transfer protein beta from Saccharomyces cerevisiae.

1991 ◽  
Vol 11 (5) ◽  
pp. 2583-2592 ◽  
Author(s):  
C C Dykstra ◽  
K Kitada ◽  
A B Clark ◽  
R K Hamatake ◽  
A Sugino
Keyword(s):  
Saccharomyces Cerevisiae ◽  
Intragenic Recombination ◽  
Temperature Sensitive ◽  
Strand Transfer ◽  
Gene Encoding ◽  
Yeast Saccharomyces Cerevisiae ◽  
Prolonged Incubation ◽  
Transfer Protein ◽  
Dna Strand

The gene encoding the 180-kDa DNA strand transfer protein beta from the yeast Saccharomyces cerevisiae was identified and sequenced. This gene, DST2 (DNA strand transferase 2), was located on chromosome VII. dst2 gene disruption mutants exhibited temperature-sensitive sporulation and a 50% longer generation time during vegetative growth than did the wild type. Spontaneous mitotic recombination in the mutants was reduced severalfold for both intrachromosomal recombination and intragenic gene conversion. The mutants also had reduced levels of the intragenic recombination that is induced during meiosis. Meiotic recombinants were, however, somewhat unstable in the mutants, with a decrease in recombinants and survival upon prolonged incubation in sporulation media. spo13 or spo13 rad50 mutations did not relieve the sporulation defect of dst2 mutations. A dst1 dst2 double mutant has the same phenotype as a dst2 single mutant. All phenotypes associated with the dst2 mutations could be complemented by a plasmid containing DST2.

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