In vivo analysis of mutated initiation codons in the mitochondrial COX2 gene of Saccharomyces cerevisiae fused to the reporter gene ARG8  m reveals lack of downstream reinitiation

2000 ◽  
Vol 262 (6) ◽  
pp. 1036-1046 ◽  
Author(s):  
N. Bonnefoy ◽  
T. D. Fox
Keyword(s):  
Saccharomyces Cerevisiae ◽  
Reporter Gene ◽  
In Vivo Analysis

scholarly journals In Vivo Analysis of Glucocorticoid-Induced Reporter Gene Expression Using Gene Gun DNA Delivery

2002 ◽  
Vol 25 (8) ◽  
pp. 1115-1118 ◽  
Author(s):  
Kiyoshi Tanigawa ◽  
Katsunao Tanaka ◽  
Hidetaka Nagase ◽  
Hidekazu Miyake ◽  
Mamoru Kiniwa ◽  
...  
Keyword(s):  
Gene Expression ◽  
Reporter Gene ◽  
Dna Delivery ◽  
Reporter Gene Expression ◽  
Gene Gun ◽  
In Vivo Analysis

Effects on mRNA splicing of mutations in the 3' region of the Saccharomyces cerevisiae actin intron

1987 ◽  
Vol 7 (1) ◽  
pp. 225-230 ◽  
Author(s):  
L A Fouser ◽  
J D Friesen
Keyword(s):  
Saccharomyces Cerevisiae ◽  
Point Mutations ◽  
Mrna Splicing ◽  
Polypyrimidine Tract ◽  
Context Change ◽  
Sequence Context ◽  
Primary Transcript ◽  
In Vivo Analysis ◽  
Specific Sequences

Point mutations, deletions, and a sequence context change were introduced at positions 3' to the internal conserved TACTAAC sequence of the Saccharomyces cerevisiae actin intron. In vivo analysis of yeast mRNA splicing suggests that, in contrast to the importance of the polypyrimidine tract in metazoan introns, specific sequences in this region are not required for efficient excision of a yeast intron. However, a double point mutation near the 3' junction (GG/AC) does severely inhibit splicing. Although this mutagenesis of the 3' junction, as well as deletion of most nucleotides between the TACTAAC and the 3' junction, caused only a slight accumulation of primary transcript, the observed accumulation of lariat intermediate by these mutants demonstrates the significance of this region for a step(s) in the splicing process after lariat formation.

scholarly journals In vivo analysis of Saccharomyces cerevisiae plasma membrane ATPase Pma1p isoforms with increased in vitro H+/ATP stoichiometry

2012 ◽  
Vol 102 (2) ◽  
pp. 401-406 ◽  
Author(s):  
Stefan de Kok ◽  
Duygu Yilmaz ◽  
Jean-Marc Daran ◽  
Jack T. Pronk ◽  
Antonius J. A. van Maris
Keyword(s):  
Saccharomyces Cerevisiae ◽  
Plasma Membrane ◽  
Plasma Membrane Atpase ◽  
Membrane Atpase ◽  
In Vivo Analysis

scholarly journals In vivo analysis of cohesin architecture using FRET in the budding yeast Saccharomyces cerevisiae

2007 ◽  
Vol 26 (16) ◽  
pp. 3783-3793 ◽  
Author(s):  
John Mc Intyre ◽  
Eric G D Muller ◽  
Stefan Weitzer ◽  
Brian E Snydsman ◽  
Trisha N Davis ◽  
...  
Keyword(s):  
Saccharomyces Cerevisiae ◽  
Budding Yeast ◽  
Yeast Saccharomyces Cerevisiae ◽  
In Vivo Analysis

scholarly journals In Vivo Analysis of Folate Coenzymes and Their Compartmentation in Saccharomyces cerevisiae

Genetics ◽  
1996 ◽  
Vol 142 (2) ◽  
pp. 371-381 ◽  
Author(s):  
J Bryan McNeil ◽  
Andrew L Bognar ◽  
Ronald E Pearlman
Keyword(s):  
Saccharomyces Cerevisiae ◽  
Serine Hydroxymethyltransferase ◽  
Mitochondrial Enzymes ◽  
Glycine Cleavage System ◽  
In Vivo Analysis ◽  
Cytoplasmic Compartment ◽  
Glycine Cleavage ◽  
In Vivo Growth ◽  
Glycine Synthesis

Abstract In eukaryotes, enzymes responsible for the interconversion of one-carbon units exist in parallel in both mitochondria and the cytoplasm. Strains of Saccharomyces cerevisiae were constructed that possess combinations of gene disruptions at the SHM1 [mitochondrial serine hydroxymethyltransferase (SHMTm)], SHM2 [cytoplasmic SHMT (SHMTc)], MIS1 [mitochondrial C1-tetrahydrofolate synthase (C1-THFSm)], ADE3 [cytoplasmic C1-THF synthase (C1-THFSc)], GCV1 [glycine cleavage system (GCV) protein T], and the GLY1 (involved in glycine synthesis) loci. Analysis of the in vivo growth characteristics and phenotypes was used to determine the contribution to cytoplasmic nucleic acid and amino acid anabolism by the mitochondrial enzymes involved in the interconversion of folate coenzymes. The data indicate that mitochondria transport formate to the cytoplasmic compartment and mitochondrial synthesis of formate appears to rely primarily on SHMTm rather than the glycine cleavage system. The glycine cleavage system and SHMTm cooperate to specifically synthesize serine. With the inactivation of SHM1, however, the glycine cleavage system can make an observable contribution to the level of mitochondrial formate. Inactivation of SHM1, SHM2 and ADE3 is required to render yeast auxotrophic for TMP and methionine, suggesting that TMP synthesized in mitochondria may be available to the cytoplasmic compartment.

scholarly journals Properties of promoters cloned randomly from the Saccharomyces cerevisiae genome.

1988 ◽  
Vol 8 (10) ◽  
pp. 4217-4224 ◽  
Author(s):  
G M Santangelo ◽  
J Tornow ◽  
C S McLaughlin ◽  
K Moldave
Keyword(s):  
Saccharomyces Cerevisiae ◽  
Rna Polymerase Ii ◽  
Reporter Gene ◽  
Reporter Genes ◽  
Rapid Identification ◽  
Base Pairs ◽  
In Vivo Assays ◽  
Tata Element ◽  
Beta Galactosidase

Promoters were isolated at random from the genome of Saccharomyces cerevisiae by using a plasmid that contains a divergently arrayed pair of promoterless reporter genes. A comprehensive library was constructed by inserting random (DNase I-generated) fragments into the intergenic region upstream from the reporter genes. Simple in vivo assays for either reporter gene product (alcohol dehydrogenase or beta-galactosidase) allowed the rapid identification of promoters from among these random fragments. Poly(dA-dT) homopolymer tracts were present in three of five randomly cloned promoters. With two exceptions, each RNA start site detected was 40 to 100 base pairs downstream from a TATA element. All of the randomly cloned promoters were capable of activating reporter gene transcription bidirectionally. Interestingly, one of the promoter fragments originated in a region of the S. cerevisiae rDNA spacer; regulated divergent transcription (presumably by RNA polymerase II) initiated in the same region.

Properties of promoters cloned randomly from the Saccharomyces cerevisiae genome

1988 ◽  
Vol 8 (10) ◽  
pp. 4217-4224
Author(s):  
G M Santangelo ◽  
J Tornow ◽  
C S McLaughlin ◽  
K Moldave
Keyword(s):  
Saccharomyces Cerevisiae ◽  
Rna Polymerase Ii ◽  
Reporter Gene ◽  
Reporter Genes ◽  
Rapid Identification ◽  
Base Pairs ◽  
In Vivo Assays ◽  
Tata Element ◽  
Beta Galactosidase

Promoters were isolated at random from the genome of Saccharomyces cerevisiae by using a plasmid that contains a divergently arrayed pair of promoterless reporter genes. A comprehensive library was constructed by inserting random (DNase I-generated) fragments into the intergenic region upstream from the reporter genes. Simple in vivo assays for either reporter gene product (alcohol dehydrogenase or beta-galactosidase) allowed the rapid identification of promoters from among these random fragments. Poly(dA-dT) homopolymer tracts were present in three of five randomly cloned promoters. With two exceptions, each RNA start site detected was 40 to 100 base pairs downstream from a TATA element. All of the randomly cloned promoters were capable of activating reporter gene transcription bidirectionally. Interestingly, one of the promoter fragments originated in a region of the S. cerevisiae rDNA spacer; regulated divergent transcription (presumably by RNA polymerase II) initiated in the same region.

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