title: Human RNase P ribonucleoprotein is required for formation of initiation complexes of RNA polymerase III.
abstract: Human RNase P is implicated in transcription of small non-coding RNA genes by RNA polymerase III (Pol III), but the precise role of this ribonucleoprotein therein remains unknown. We here show that targeted destruction of HeLa nuclear RNase P inhibits transcription of 5S rRNA genes in whole cell extracts, if this precedes the stage of initiation complex formation. Biochemical purification analyses further reveal that this ribonucleoprotein is recruited to 5S rRNA genes as a part of proficient initiation complexes and the activity persists at reinitiation. Knockdown of RNase P abolishes the assembly of initiation complexes by preventing the formation of the initiation sub-complex of Pol III. Our results demonstrate that the structural intactness, but not the endoribonucleolytic activity per se, of RNase P is critical for the function of Pol III in cells and in extracts.
authors.affiliation: Department of Microbiology and Molecular Genetics, IMRIC, The Hebrew University-Hadassah Medical School, Jerusalem 91120, Israel.
authors.forename: Raphael
authors.initials: R
authors.lastname: Serruya
authors.affiliation: Department of Microbiology and Molecular Genetics, IMRIC, The Hebrew University-Hadassah Medical School, Jerusalem 91120, Israel.
authors.forename: Natalie
authors.initials: N
authors.lastname: Orlovetskie
authors.affiliation: Department of Microbiology and Molecular Genetics, IMRIC, The Hebrew University-Hadassah Medical School, Jerusalem 91120, Israel.
authors.forename: Robert
authors.initials: R
authors.lastname: Reiner
authors.affiliation: Department of Microbiology and Molecular Genetics, IMRIC, The Hebrew University-Hadassah Medical School, Jerusalem 91120, Israel.
authors.forename: Yana
authors.initials: Y
authors.lastname: Dehtiar-Zilber
authors.affiliation: Department of Molecular, Cellular and Developmental Biology, Yale University, New Haven, CT 06520, USA.
authors.forename: Donna
authors.initials: D
authors.lastname: Wesolowski
authors.affiliation: Department of Molecular, Cellular and Developmental Biology, Yale University, New Haven, CT 06520, USA.
authors.forename: Sidney
authors.initials: S
authors.lastname: Altman
authors.affiliation: Department of Microbiology and Molecular Genetics, IMRIC, The Hebrew University-Hadassah Medical School, Jerusalem 91120, Israel jarrous@md.huji.ac.il.
authors.forename: Nayef
authors.initials: N
authors.lastname: Jarrous
mesh_terms: D006367:HeLa Cells; D006801:Humans; D012320:RNA Polymerase III / Q000378:metabolism*; D012341:RNA, Ribosomal, 5S / Q000235:genetics*; D022661:RNA, Untranslated / Q000378:metabolism; D043262:Ribonuclease P / Q000037:antagonists & inhibitors / Q000302:isolation & purification / Q000378:metabolism*; D061785:Transcription Initiation, Genetic
chemical_list: D012341:RNA, Ribosomal, 5S; D022661:RNA, Untranslated; D012320:RNA Polymerase III; D043262:Ribonuclease P