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Nucleic Acids Research, 1979, Vol. 6, No. 5 1929-1940
© 1979

Articles

Properties of in vitro trarscription by isolated Xenopus oocyte nucleoli

Hidctoshi saiga and Toru Higashinakagawa

Laboratory of Developmental Biology, Mitsubishi-Kasei Institute of Life Sciences, 11 Minamiooya Machida, Tokyo, Japan

Received March 8, 1979. Some properties of in vitro transcription by isolated Xenopus oocyte nucleoli were described. When incubated with labeled ERA precursors, Xenopus oocyte nucleoli exhibited prolonged incorporation of radioactivity into BRA. The synthetic activity was exclusively due to type I ERA polymerase as revealed by its insensitivity to low and high doses of {alpha}-ananitin. The size of the in vitro transcript was mostly larger than 28S at 10 minute incubation and became smaller as incubation proceeded. When [{gamma}-32p] ATP was included in the reaction mixture, 32 radioactivity was incorporated into BRA suggesting the possible initiation of transcription in this system. However, analysis of the terminal nucleotide of the transcript revealed that the incorporation of radioactivity from [{gamma}-32p] was not due to the initiation of transcrip tion but due to polynucleotide kinase activity in the nucleolar preparation. These results demonstrate that the incorporation of radioactivity from [{gamma}-32p] labeled nucleoside triphosphates cannot necessarily be regarded as an index of the initiation of transcription.


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