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pubmed-article:10982336pubmed:abstractTextBaculovirus late RNAs are transcribed by a four-subunit RNA polymerase that is virus encoded. The late viral mRNAs are capped and polyadenylated, and we have previously shown that capping is mediated by the LEF-4 subunit of baculovirus RNA polymerase. Here we report studies undertaken to determine the mechanism of 3'-end formation. A globin cleavage/polyadenylation signal, which was previously shown to direct 3'-end formation of viral RNAs in vivo, was cloned into a baculovirus transcription template. In vitro assays with purified baculovirus RNA polymerase revealed that 3' ends were formed not by a cleavage mechanism but rather by termination after transcription of a T-rich region of the globin sequence. Terminated RNAs were released from ternary complexes and were subsequently polyadenylated. Mutational analyses indicated that the T-rich sequence was essential for termination and polyadenylation, but the poly(A) signal and the GT-rich region of the globin polyadenylation/cleavage signal were not required. Termination was not dependent on ATP hydrolysis, indicating a slippage mechanism.lld:pubmed
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pubmed-article:10982336pubmed:dateRevised2009-11-18lld:pubmed
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pubmed-article:10982336pubmed:articleTitle3'-end formation of baculovirus late RNAs.lld:pubmed
pubmed-article:10982336pubmed:affiliationDepartment of Biochemistry and Biophysics, Texas A&M University, College Station, Texas 77843-2128, USA.lld:pubmed
pubmed-article:10982336pubmed:publicationTypeJournal Articlelld:pubmed
pubmed-article:10982336pubmed:publicationTypeResearch Support, U.S. Gov't, Non-P.H.S.lld:pubmed
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