title: Identification and characterization of a nuclease specific for the 3' end of the U6 small nuclear RNA.
abstract: The U6 small nuclear RNA is post-transcriptionally processed by the addition and removal of nontemplate uridylates (Us) at its 3' end prior to incorporation into the U4 x U6 small nuclear ribonucleoprotein complex. An enzyme responsible for removing Us from the U6 3' end has not been previously identified. Here we biochemically isolate and characterize an exonuclease activity from HeLa cells that removes template and nontemplate 3'-nucleotides specifically from U6 RNA. We also report the isolation of an inhibitor of this U6 nuclease. U6 nuclease rapidly removes the four terminal 3' Us found in the human U6 coding sequence in a magnesium-dependent manner. Mutagenesis studies on the U6 RNA define regions essential for processing. U6 nuclease recognizes specific sequences on both strands near the base of the major intramolecular stem loop of the U6 RNA. The preprocessed 3' Us form part of the base of the stem loop, but neither specific sequences nor secondary structure at the four terminal nucleotides are required to achieve processing.
authors.affiliation: Department of Biochemistry and Molecular Biology, The Pennsylvania State University, University Park 16802, USA.
authors.forename: B L
authors.initials: BL
authors.lastname: Booth
authors.forename: B F
authors.initials: BF
authors.lastname: Pugh
mesh_terms: D001483:Base Sequence; D005095:Exoribonucleases / Q000737:chemistry / Q000378:metabolism*; D006801:Humans; D007700:Kinetics; D008274:Magnesium / Q000378:metabolism; D008969:Molecular Sequence Data; D016296:Mutagenesis; D009690:Nucleic Acid Conformation; D011499:Protein Processing, Post-Translational; D012342:RNA, Small Nuclear / Q000378:metabolism*; D017384:Sequence Deletion
chemical_list: D012342:RNA, Small Nuclear; D005095:Exoribonucleases; C103661:U6 nuclease; D008274:Magnesium