Eukaryotes Have Three Nuclear RNA Polymerases

Unlike prokaryotes, in which all major types of RNA are synthesized by one RNA polymerase, eukaryotic cells contain three nuclear DNA-dependent RNA polymerases, each responsible for synthesizing a different class of RNAs. 

Nuclear extracts can be fractionated by chromatography on DEAE-cellulose to give three peaks of RNA polymerase activity (the use of column chromatography is explained in chapter 6). These three peaks correspond to three different RNA polymerases (I, II, and III), which differ in relative amount, cellular location, type of RNA synthesized, subunit structure, response to salt and divalent cation concentrations, and sensitivity to the mushroom-derived toxin a-amanitin. The three polymerases and some of their properties are summarized in table 28.4. 

RNA polymerase I is located in the nucleolus and synthesizes a large precursor that is later processed to form rRNA. It is completely resistant to inhibition by a-amanitin. RNA polymerase II is located in the nucleoplasm and synthesizes large precursor RNAs (sometimes called heterogeneous nuclear RNA, or hnRNA) that are processed to form cytoplasmic mRNAs. It is also responsible for the synthesis of most viral RNA in virus-infected cells. PolII is very sensitive to a-amanitin, being inhibited by 50% at 0.05 /u,g/ml. RNA polymerase III is also located in the nucleoplasm and synthesizes small RNAs, such as 5S RNA and the precursors to tRNAs. This enzyme is somewhat resistant to a-amanitin, requiring about 5 /u,g/ml to reach 50% inhibition. 

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Eukaryotes Have Three Nuclear RNA Polymerases Eukaryotic RNA Polymerases Are Not Fully Functional by Themselves... 

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