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Predicate | Object |
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rdf:type | |
lifeskim:mentions | |
pubmed:issue |
2
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pubmed:dateCreated |
1992-1-17
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pubmed:abstractText |
A human eIF-2 alpha cDNA (encoding alpha-subunit of the eukaryotic initiation factor-2) was expressed under the control of the galactose-regulated GAL1, 10 promoter, in Saccharomyces cerevisiae, in order to study the possible interactions of human eIF-2 alpha with the yeast protein synthesis apparatus. Isoelectric focusing coupled with Western-blot analysis demonstrated that the human eIF-2 alpha subunit synthesized in yeast under a variety of growth conditions was detected as two bands which co-migrated with the phosphorylated and unphosphorylated forms of rabbit eIF-2 alpha, suggesting covalent modification in vivo. Cell fractionation studies further demonstrated that the synthesised human eIF-2 alpha protein, though present in the cytoplasm, was largely associated with the yeast ribosomes, but could be removed from these by washing with 0.3 M KCl. This possible association of the synthesised human subunit into a three-subunit (alpha, beta and gamma) eIF-2 complex was further examined by partial purification of the yeast eIF-2 complex and estimation of the molecular mass of this complex. Immunoreactive eIF-2 alpha was found in fractions with eIF-2 activity and the estimated molecular mass (130 kDa) corresponded to that predicted for the eIF-2 trimer. These analyses suggest that human eIF-2 alpha subunit synthesised in yeast can become involved with the yeast protein synthetic apparatus, though whether this is a functional incorporation requires further genetic studies.
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pubmed:language |
eng
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pubmed:journal | |
pubmed:citationSubset |
IM
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pubmed:chemical | |
pubmed:status |
MEDLINE
|
pubmed:month |
Dec
|
pubmed:issn |
0378-1119
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pubmed:author | |
pubmed:issnType |
Print
|
pubmed:day |
15
|
pubmed:volume |
108
|
pubmed:owner |
NLM
|
pubmed:authorsComplete |
Y
|
pubmed:pagination |
253-8
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pubmed:dateRevised |
2006-11-15
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pubmed:meshHeading |
pubmed-meshheading:1748310-Eukaryotic Initiation Factor-2,
pubmed-meshheading:1748310-Galactose,
pubmed-meshheading:1748310-Gene Expression Regulation, Fungal,
pubmed-meshheading:1748310-Humans,
pubmed-meshheading:1748310-Plasmids,
pubmed-meshheading:1748310-Protein Processing, Post-Translational,
pubmed-meshheading:1748310-Recombinant Proteins,
pubmed-meshheading:1748310-Saccharomyces cerevisiae,
pubmed-meshheading:1748310-Transformation, Genetic
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pubmed:year |
1991
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pubmed:articleTitle |
Synthesis of human initiation factor-2 alpha in Saccharomyces cerevisiae.
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pubmed:affiliation |
Biological Laboratory, University of Kent, Canterbury, U.K.
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pubmed:publicationType |
Journal Article,
Research Support, Non-U.S. Gov't
|