Open AccessTranslation in vitro of rat brain messenger RNA coding for tubulin and actin.
Author(s) -
Illana Gozes,
Henri Schmitt,
Uriel Z. Littauer
Publication year - 1975
Publication title -
proceedings of the national academy of sciences of the united states of america
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 5.011
H-Index - 771
eISSN - 1091-6490
pISSN - 0027-8424
DOI - 10.1073/pnas.72.2.701
Subject(s) - tubulin , sodium dodecyl sulfate , actin , biochemistry , gel electrophoresis , polyacrylamide gel electrophoresis , in vitro , colchicine , microtubule , messenger rna , urea , rna , chemistry , cytoskeleton , biology , microbiology and biotechnology , cell , enzyme , genetics , gene
A partially purified fraction of poly(a)-rich brain mRNA coding for tubulin and actin was obtained by stepwise elution from an oligo(deoxythymidylate)-cellulose column and was efficiently translated in a wheat-germ cell-free system. The newly synthesized tubulin and actin migrated along with the authentic proteins on sodium dodecyl sulfate polyacrylamide gels and on sodium dodecyl sulfate-urea polyacrylamide gels, where tubulin alpha- and beta-subunits are separated. The two proteins synthesized in vitro were found to be biologically active; they could be induced to polymerize and were both precipitated by vinblastine. In addition, specific binding of tubulin to an affinity column of colchicine-Sepharose and actin to myosin were demonstrated.