Transformation of Intact Yeast Cells Treated with Alkali Cations or Thiol Compounds

Ito, Hisao; Murata, Kousaku; Kimura, Akira

doi:10.1080/00021369.1984.10866161

Cited by 416 publications

(438 citation statements)

References 3 publications

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“…Yeast plasmids and linear DNA fragments for integration were transformed into yeast by lithium acetate transformation (Ito et al, 1983). Selection against Ura+ strains was done by culture on solid synthetic media containing 1 mg/ml 5-fluoroorotic acid (5-FOA) as described in Boeke et al (1984).…”

Section: Yeast Growth Conditions and Strain Constructionmentioning

confidence: 99%

NUP2, a novel yeast nucleoporin, has functional overlap with other proteins of the nuclear pore complex.

Loeb

Davis

Fink

1993

MBoC

116

125

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We have isolated a new gene, NUP2, that encodes a constituent of the yeast-nuclear pore complex (NPC). The NUP2 protein sequence shares a central repetitive domain with NSP1 and NUP1, the two previously characterized yeast nucleoporins. Like NUP1 and NSP1, NUP2 localizes to discrete spots in the nuclear envelope, as determined by indirect immunofluorescence. Although the sequence similarity among these three nucleoporins suggests that they have a similar role in the nuclear pore complex, NUP2, in contrast to NSP1 and NUP1, is not required for growth. Some combinations of mutant alleles of NUP1, NSP1, and NUP2 display "synthetic lethal" relationships that provide evidence for functional interaction between these NPC components. This genetic evidence of overlapping function suggests that the nucleoporins act in concert, perhaps participating in the same step of the recognition or transit of macromolecules through the NPC.

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Section: Yeast Growth Conditions and Strain Constructionmentioning

confidence: 99%

NUP2, a novel yeast nucleoporin, has functional overlap with other proteins of the nuclear pore complex.

Loeb

Davis

Fink

1993

MBoC

116

125

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“…Strains H13-3774 and 314L-3914 were generated by introducing plasmids encoding Hcr1p or Rpg1p (courtesy of Alan Hinnebusch, NIH) by standard cation transformation. 69 Strains H2545 (MATa IMT1 IMT2 imt3::TRP1 imt4::TRP1 trp1-GAL + ura3-52 i leu2::HISG), H2546 (MATa IMT1 IMT2 imt3::TRP1 imt4::TRP1 trp1-GAL + ura3-52 i leu2::HISG gcn2Δ), H4 (MATa leu2-3,112 ura3-52), H952 dependent initiation. The reason for the different findings in yeast and mammalian cells is not known.…”

Section: Methodsmentioning

confidence: 99%

Components of the multifactor complex needed for internal initiation by the IRES of hepatitis C virus inSaccharomyces cerevisiae

Rosenfeld

Racaniello

2010

RNA Biology

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Interaction between the 40S ribosomal subunit and the IRES of hepatitis C virus (HCV) is thought to be independent of initiation proteins, while joining of the 60S ribosomal subunit, and initiation of translation is dependent upon components of the translation machinery. An established in vivo functional assay for internal initiation mediated by the HCV IRES was used to identify proteins needed for IRES dependent translation in Saccharomyces cerevisiae strains possessing alterations of the translation machinery. Internal initiation dependent upon the HCV IRES was abrogated in strains lacking eIF5B, and reduced in strains with altered eIF3, either lacking the Hcr1p subunit, a component of eIF3 not previously known to interact with HCV RNA, or possessing an amino acid change in the Rpg1p subunit. The HCV RNA-induced conformational change in the 40S subunit might affect positioning of eIF3 and lead to different interactions between the ribosome, eIF3, and the multifactor complex. HCV IRES dependent initiation was unaffected in strains in which the concentration of the initiating tRNA was reduced. Alteration of the δ subunit of eIF2B, which leads to inefficient recycling, or substitution of aspartic acid for serine 51 of eIF2α had no effect on internal initiation. Production of human Pkr inhibited HCV IRES dependent initiation in yeast. The synthesis of Pkr in yeast is known to result in high levels of eIF2α phosphorylation, increased Gcn4p synthesis, and reduced ribosomal protein production. These alterations may explain the effect of Pkr synthesis on HCV IRES dependent initiation in yeast.

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“…Among the lithium salts tested, LiCI and LiN0 3 were effective to attain high transformation efficiencies. Lithium acetate, which was highly effective in the induction of competence in S. cerevisiae, 23) was not as effective in this strain. Optimization of the transformation condition was done using intact cells treated with LiN0 3 • The optimum concentration and the duration of LiN0 3 treatment were roughly between 200 and 600 mM (Fig.…”

Section: Transformation Of K Thermotolerans Using the U Ra3 Gene Of mentioning

confidence: 99%

“…Among the thiol compounds tested, 2-mercaptoethanol, 3-mercapto-l,2-propanediol and 2-mercaptoethylamine were effective as in the case of S. cerevisiae. 23 ) However, the effects of these compounds on transformation frequency varied widely from one experiment to another, and it appeared that the stimulatory effects of these compounds decreased with the increase of the period of storage of the reagents (data not shown).…”

Section: Transformation Of K Thermotolerans Using the U Ra3 Gene Of mentioning

confidence: 99%

Construction of a Genetic Transformation System for a Freeze-tolerant YeastKluyveromyces thermotolerans

Hino

Wongkhalaung

Kawai

et al. 1992

Bioscience, Biotechnology, and Biochemistry

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Transformation of Intact Yeast Cells Treated with Alkali Cations or Thiol Compounds

Cited by 416 publications

References 3 publications

NUP2, a novel yeast nucleoporin, has functional overlap with other proteins of the nuclear pore complex.

NUP2, a novel yeast nucleoporin, has functional overlap with other proteins of the nuclear pore complex.

Components of the multifactor complex needed for internal initiation by the IRES of hepatitis C virus inSaccharomyces cerevisiae

Construction of a Genetic Transformation System for a Freeze-tolerant YeastKluyveromyces thermotolerans

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