title: The NIP7 protein is required for accurate pre-rRNA processing in human cells.
abstract: Eukaryotic ribosome biogenesis requires the function of a large number of trans-acting factors which interact transiently with the nascent pre-rRNA and dissociate as the ribosomal subunits proceed to maturation and export to the cytoplasm. Loss-of-function mutations in human trans-acting factors or ribosome components may lead to genetic syndromes. In a previous study, we have shown association between the SBDS (Shwachman-Bodian-Diamond syndrome) and NIP7 proteins and that downregulation of SBDS in HEK293 affects gene expression at the transcriptional and translational levels. In this study, we show that downregulation of NIP7 affects pre-rRNA processing, causing an imbalance of the 40S/60S subunit ratio. We also identified defects at the pre-rRNA processing level with a decrease of the 34S pre-rRNA concentration and an increase of the 26S and 21S pre-rRNA concentrations, indicating that processing at site 2 is particularly slower in NIP7-depleted cells and showing that NIP7 is required for maturation of the 18S rRNA. The NIP7 protein is restricted to the nuclear compartment and co-sediments with complexes with molecular masses in the range of 40S-80S, suggesting an association to nucleolar pre-ribosomal particles. Downregulation of NIP7 affects cell proliferation, consistently with an important role for NIP7 in rRNA biosynthesis in human cells.
authors.affiliation: Laboratório Nacional de Biociências, Centro Nacional de Pesquisa em Energia e Materiais, Campinas SP, Brazil.
authors.forename: Luis G
authors.initials: LG
authors.lastname: Morello
authors.forename: Cédric
authors.initials: C
authors.lastname: Hesling
authors.forename: Patrícia P
authors.initials: PP
authors.lastname: Coltri
authors.forename: Beatriz A
authors.initials: BA
authors.lastname: Castilho
authors.forename: Ruth
authors.initials: R
authors.lastname: Rimokh
authors.forename: Nilson I T
authors.initials: NI
authors.lastname: Zanchin
mesh_terms: D002460:Cell Line; D022003:Cell Nucleus Structures / Q000737:chemistry; D055785:Gene Knockdown Techniques; D057809:HEK293 Cells; D006801:Humans; D009687:Nuclear Proteins / Q000037:antagonists & inhibitors / Q000235:genetics / Q000502:physiology*; D011063:Poly A-U / Q000378:metabolism; D011072:Poly U / Q000378:metabolism; D011132:Polyribosomes / Q000737:chemistry; D012313:RNA / Q000737:chemistry / Q000378:metabolism; D012322:RNA Precursors / Q000737:chemistry / Q000378:metabolism*; D012323:RNA Processing, Post-Transcriptional; D012335:RNA, Ribosomal / Q000737:chemistry / Q000378:metabolism*
chemical_list: C499553:NIP7 protein, human; D009687:Nuclear Proteins; D012322:RNA Precursors; D012335:RNA, Ribosomal; D011063:Poly A-U; D011072:Poly U; D012313:RNA
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