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Proper 3' end formation of the human pre-snRNAs synthesized by pol II requires the cis-acting 3' box, although the precise function of this element has proved difficult to determine. In vivo, 3' end formation is tightly linked to transcription. However, we have now been able to obtain transcription-independent 3' box-dependent processing in vitro. This finally demonstrates that the 3' end of pre-snRNAs is produced by RNA processing rather than by termination of transcription. The phosphorylated form of the C-terminal domain (CTD) of pol II activates the processing event in vitro, consistent with our previous demonstration of the role of the CTD in pre-snRNA 3' end formation in vivo. In addition, we show that sequences upstream from the 3' box of the U2 snRNA gene influence 3' end formation both in vivo and in vitro.

Original publication

DOI

10.1093/emboj/cdg430

Type

Journal article

Journal

EMBO J

Publication Date

01/09/2003

Volume

22

Pages

4544 - 4554

Keywords

Base Sequence, Cations, Divalent, DNA, HeLa Cells, Humans, In Vitro Techniques, Models, Biological, Nucleic Acid Conformation, Protein Structure, Tertiary, RNA Polymerase II, RNA Precursors, RNA Processing, Post-Transcriptional, RNA, Small Nuclear, Recombinant Fusion Proteins, Transcription, Genetic