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PredicateObject
rdf:type
lifeskim:mentions
pubmed:issue
18
pubmed:dateCreated
1976-11-21
pubmed:abstractText
Experimental evidence showing specificity of rabbit reticulocyte initiation factor 3 (EIF-3) for selective initiation of mRNA translation is presented. A new cell-free system was developed from Crithidia fasciculata. The crude postmitochondrial supernatant fluid was treated with puromycin and 0.5 M KCl to dissociate mRNA from polysomes and ribosomes into subunits. The drug and salt were removed by gel filtration on Sephadex G-25. Additions of amino acids and energy source initiate protein synthesis. All synthesis starts at the initiation site. This treatment brought about a shift in MgCl2 optimum from 6 to 3 mM. Exogenously supplied rabbit reticulocyte globin mRNA is faithfully translated in this system. However, crithidial EIF-3 has a low affinity for globin mRNA as evidenced by a 6-fold increase in the rate of globin synthesis after the addition of rabbit reticulocyte EIF-3 in the range at which globin synthesis is linear to the amount of globin mRNA added to the system. It is also shown that in a reconstituted system in which ribosomal subunits are depleted from initiation factors, EIF-3 from rabbit reticulocytes has a higher affinity for globin mRNA, as measured by the formation of polysomes during the linear time of amino acid incorporation. These results are taken to indicate that initiation factor EIF-3 action should be considered as an enzyme catalyzed reaction for which various mRNAs serve as different substrate analogs. Therefore, specificity is most likely to be expressed as an affinity of enzyme to substrate and would show as rate difference rather than an all-or-none phenomenon.
pubmed:language
eng
pubmed:journal
pubmed:citationSubset
IM
pubmed:chemical
pubmed:status
MEDLINE
pubmed:month
Sep
pubmed:issn
0021-9258
pubmed:author
pubmed:issnType
Print
pubmed:day
25
pubmed:volume
251
pubmed:owner
NLM
pubmed:authorsComplete
Y
pubmed:pagination
5718-25
pubmed:dateRevised
2009-11-19
pubmed:meshHeading
pubmed:year
1976
pubmed:articleTitle
Requirement for homologous rabbit reticulocyte initiation factor 3 for initiation of alpha- and beta-globin mRNA translation in a crude protozoal cell-free system.
pubmed:publicationType
Journal Article, Research Support, U.S. Gov't, P.H.S.