title: RBM7 subunit of the NEXT complex binds U-rich sequences and targets 3'-end extended forms of snRNAs.
abstract: The Nuclear Exosome Targeting (NEXT) complex is a key cofactor of the mammalian nuclear exosome in the removal of Promoter Upstream Transcripts (PROMPTs) and potentially aberrant forms of other noncoding RNAs, such as snRNAs. NEXT is composed of three subunits SKIV2L2, ZCCHC8 and RBM7. We have recently identified the NEXT complex in our screen for oligo(U) RNA-binding factors. Here, we demonstrate that NEXT displays preference for U-rich pyrimidine sequences and this RNA binding is mediated by the RNA recognition motif (RRM) of the RBM7 subunit. We solved the structure of RBM7 RRM and identified two phenylalanine residues that are critical for interaction with RNA. Furthermore, we showed that these residues are required for the NEXT interaction with snRNAs in vivo. Finally, we show that depletion of components of the NEXT complex alone or together with exosome nucleases resulted in the accumulation of mature as well as extended forms of snRNAs. Thus, our data suggest a new scenario in which the NEXT complex is involved in the surveillance of snRNAs and/or biogenesis of snRNPs.
authors: CEITEC-Central European Institute of Technology, Masaryk University, Brno, 62500, Czech Republic National Centre for Biomolecular Research, Faculty of Science, Masaryk University, Brno, 62500, Czech Republic.
authors: Dominika
authors: D
authors: Hrossova
authors: CEITEC-Central European Institute of Technology, Masaryk University, Brno, 62500, Czech Republic National Centre for Biomolecular Research, Faculty of Science, Masaryk University, Brno, 62500, Czech Republic.
authors: Tomas
authors: T
authors: Sikorsky
authors: CEITEC-Central European Institute of Technology, Masaryk University, Brno, 62500, Czech Republic.
authors: David
authors: D
authors: Potesil
authors: CEITEC-Central European Institute of Technology, Masaryk University, Brno, 62500, Czech Republic National Centre for Biomolecular Research, Faculty of Science, Masaryk University, Brno, 62500, Czech Republic.
authors: Marek
authors: M
authors: Bartosovic
authors: CEITEC-Central European Institute of Technology, Masaryk University, Brno, 62500, Czech Republic.
authors: Josef
authors: J
authors: Pasulka
authors: CEITEC-Central European Institute of Technology, Masaryk University, Brno, 62500, Czech Republic.
authors: Zbynek
authors: Z
authors: Zdrahal
authors: CEITEC-Central European Institute of Technology, Masaryk University, Brno, 62500, Czech Republic National Centre for Biomolecular Research, Faculty of Science, Masaryk University, Brno, 62500, Czech Republic richard.stefl@ceitec.muni.cz.
authors: Richard
authors: R
authors: Stefl
authors: CEITEC-Central European Institute of Technology, Masaryk University, Brno, 62500, Czech Republic vanacova@chemi.muni.cz.
authors: Stepanka
authors: S
authors: Vanacova
mesh_terms: D020816:Amino Acid Motifs; D001483:Base Sequence; D057809:HEK293 Cells; D006367:HeLa Cells; D006801:Humans; D009843:Oligoribonucleotides / Q000378:metabolism; D011485:Protein Binding; D021122:Protein Subunits / Q000737:chemistry / Q000378:metabolism; D012342:RNA, Small Nuclear / Q000737:chemistry / Q000378:metabolism*; D016601:RNA-Binding Proteins / Q000032:analysis / Q000737:chemistry* / Q000378:metabolism*; D014500:Uracil Nucleotides / Q000378:metabolism
chemical_list: D009843:Oligoribonucleotides; D021122:Protein Subunits; C000596172:RBM7 protein, human; D012342:RNA, Small Nuclear; D016601:RNA-Binding Proteins; D014500:Uracil Nucleotides; C027916:oligo(U)