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Cell ; 117(3): 311-21, 2004 Apr 30.
Article in English | MEDLINE | ID: mdl-15109492

ABSTRACT

tRNA splicing is a fundamental process required for cell growth and division. The first step in tRNA splicing is the removal of introns catalyzed in yeast by the tRNA splicing endonuclease. The enzyme responsible for intron removal in mammalian cells is unknown. We present the identification and characterization of the human tRNA splicing endonuclease. This enzyme consists of HsSen2, HsSen34, HsSen15, and HsSen54, homologs of the yeast tRNA endonuclease subunits. Additionally, we identified an alternatively spliced isoform of SEN2 that is part of a complex with unique RNA endonuclease activity. Surprisingly, both human endonuclease complexes are associated with pre-mRNA 3' end processing factors. Furthermore, siRNA-mediated depletion of SEN2 exhibited defects in maturation of both pre-tRNA and pre-mRNA. These findings demonstrate a link between pre-tRNA splicing and pre-mRNA 3' end formation, suggesting that the endonuclease subunits function in multiple RNA-processing events.


Subject(s)
Endonucleases/chemistry , Endonucleases/metabolism , RNA Precursors/metabolism , RNA Splicing , RNA, Transfer/metabolism , Amino Acid Sequence , Cell Line , Cell Nucleolus/chemistry , Cell Nucleolus/metabolism , Cell Nucleus/chemistry , Cell Nucleus/metabolism , Consensus Sequence , Conserved Sequence , Endonucleases/isolation & purification , Endoribonucleases/metabolism , HeLa Cells , Humans , Introns , Isoenzymes/chemistry , Isoenzymes/genetics , Isoenzymes/metabolism , Molecular Sequence Data , Protein Subunits , RNA Precursors/genetics , RNA Processing, Post-Transcriptional , RNA, Small Interfering/metabolism , RNA, Transfer/chemistry , Saccharomyces cerevisiae/enzymology , Sequence Homology, Amino Acid
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