Characterization and Translation of Poly(A)^+RNA from Wounded and Crown Gall Tissues of Potato Tubers

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Poly(A)^+ and poly(A)^-RNA from wounded potato tuber tissues and crown gall tumors were separated from total RNA by oligodeoxythymidylic acid-cellulose affinity chromatography. The poly(A)^+RNA was characterized by sucrose density gradient centrifugation, hybridization with ^3(H)polyuridylic acid [Poly(U)] and in vitro translation in a rabbit reticulocyte lysate system. The tumor poly(A)^+RNA was a heterodisperse mixture from 3.5S to 35S. Upon poly(U) hybridization of the gradient fractions two major hybridization peaks at 7S and 21S and two peaks at 11S and 16S appeared. In an in vitro translation system the poly(A)^+RNA programmed the synthesis of 23 different polypeptides of 9,000 to 79,800 daltons molecular weight as determined by SDS-polyacrylamide gel electrophoresis. The 21S poly(A)^+RNA was about 5 times more active in in vitro protein synthesis than the 7S poly(A)^+RNA. The poly(A)^+RNA from wounded tissues was also heterodisperse (from 4.5S to 31S) with a modal peak at 18S. This RNA coded for at least 28 polypeptides, which were different from those of crown gall tumor tissues. On a per unit poly(A)^+RNA basis the tumor RNA was slightly more active in translation than that from wounded tissues. The translation of tumor poly(A)^+RNA was completely blocked by 0.5mM 7-methylguanosine 5'-phosphate, but not by 7-methylguanosine, suggesting the presence of a 5'-cap structure.
日本植物生理学会の論文