RNA mimicry by the fap7 adenylate kinase in ribosome biogenesis.
During biogenesis of the 40S and 60S ribosomal subunits, the pre-40S particles are exported to the cytoplasm prior to final cleavage of the 20S pre-rRNA to mature 18S rRNA. Amongst the factors involved in this maturation step, Fap7 is unusual, as it both interacts with ribosomal protein Rps14 and harbors adenylate kinase activity, a function not usually associated with ribonucleoprotein assembly. Human hFap7 also regulates Cajal body assembly and cell cycle progression via the p53-MDM2 pathway. This work presents the functional and structural characterization of the Fap7-Rps14 complex. We report that Fap7 association blocks the RNA binding surface of Rps14 and, conversely, Rps14 binding inhibits adenylate kinase activity of Fap7. In addition, the affinity of Fap7 for Rps14 is higher with bound ADP, whereas ATP hydrolysis dissociates the complex. These results suggest that Fap7 chaperones Rps14 assembly into pre-40S particles via RNA mimicry in an ATP-dependent manner. Incorporation of Rps14 by Fap7 leads to a structural rearrangement of the platform domain necessary for the pre-rRNA to acquire a cleavage competent conformation.
Laboratoire de Cristallographie et RMN Biologiques, UMR CNRS 8015, Université Paris Descartes, Faculté de Pharmacie, Sorbonne Paris Cité, Paris, France.
Jérôme
J
Loc'h
Laboratoire de Cristallographie et RMN Biologiques, UMR CNRS 8015, Université Paris Descartes, Faculté de Pharmacie, Sorbonne Paris Cité, Paris, France.
Magali
M
Blaud
Laboratoire de Cristallographie et RMN Biologiques, UMR CNRS 8015, Université Paris Descartes, Faculté de Pharmacie, Sorbonne Paris Cité, Paris, France.
Stéphane
S
Réty
Laboratoire de Cristallographie et RMN Biologiques, UMR CNRS 8015, Université Paris Descartes, Faculté de Pharmacie, Sorbonne Paris Cité, Paris, France; Wellcome Trust Centre for Cell Biology, University of Edinburgh, Edinburgh, Scotland.
Simon
S
Lebaron
Laboratoire de Cristallographie et RMN Biologiques, UMR CNRS 8015, Université Paris Descartes, Faculté de Pharmacie, Sorbonne Paris Cité, Paris, France.
Patrick
P
Deschamps
Laboratoire de Cristallographie et RMN Biologiques, UMR CNRS 8015, Université Paris Descartes, Faculté de Pharmacie, Sorbonne Paris Cité, Paris, France.
Joseph
J
Bareille
Laboratoire de Cristallographie et RMN Biologiques, UMR CNRS 8015, Université Paris Descartes, Faculté de Pharmacie, Sorbonne Paris Cité, Paris, France.
Julie
J
Jombart
Laboratoire de Cristallographie et RMN Biologiques, UMR CNRS 8015, Université Paris Descartes, Faculté de Pharmacie, Sorbonne Paris Cité, Paris, France.
Julien
J
Robert-Paganin
Laboratoire de Cristallographie et RMN Biologiques, UMR CNRS 8015, Université Paris Descartes, Faculté de Pharmacie, Sorbonne Paris Cité, Paris, France.
Lila
L
Delbos
Laboratoire de Cristallographie et RMN Biologiques, UMR CNRS 8015, Université Paris Descartes, Faculté de Pharmacie, Sorbonne Paris Cité, Paris, France.
Florian
F
Chardon
Laboratoire de Cristallographie et RMN Biologiques, UMR CNRS 8015, Université Paris Descartes, Faculté de Pharmacie, Sorbonne Paris Cité, Paris, France.
Elodie
E
Zhang
Laboratoire de Cristallographie et RMN Biologiques, UMR CNRS 8015, Université Paris Descartes, Faculté de Pharmacie, Sorbonne Paris Cité, Paris, France.
Clément
C
Charenton
Wellcome Trust Centre for Cell Biology, University of Edinburgh, Edinburgh, Scotland.
David
D
Tollervey
Laboratoire de Cristallographie et RMN Biologiques, UMR CNRS 8015, Université Paris Descartes, Faculté de Pharmacie, Sorbonne Paris Cité, Paris, France.
Nicolas
N
Leulliot
D000244:Adenosine Diphosphate / Q000737:chemistry / Q000378:metabolism; D000255:Adenosine Triphosphate / Q000737:chemistry / Q000378:metabolism; D000263:Adenylate Kinase / Q000737:chemistry / Q000235:genetics* / Q000378:metabolism; D000595:Amino Acid Sequence; D004926:Escherichia coli / Q000235:genetics / Q000378:metabolism; D015966:Gene Expression Regulation, Fungal; D006801:Humans; D008958:Models, Molecular; D018716:Molecular Mimicry; D008969:Molecular Sequence Data; D009687:Nuclear Proteins / Q000737:chemistry / Q000235:genetics* / Q000378:metabolism; D043583:Nucleoside-Triphosphatase / Q000737:chemistry / Q000235:genetics* / Q000378:metabolism; D043903:Pyrococcus abyssi / Q000235:genetics / Q000378:metabolism; D012337:RNA, Ribosomal, 18S / Q000737:chemistry / Q000235:genetics / Q000378:metabolism; D011994:Recombinant Proteins / Q000737:chemistry / Q000235:genetics / Q000378:metabolism; D012269:Ribosomal Proteins / Q000737:chemistry / Q000235:genetics* / Q000378:metabolism; D054683:Ribosome Subunits, Large, Eukaryotic / Q000235:genetics / Q000378:metabolism; D054682:Ribosome Subunits, Small, Eukaryotic / Q000235:genetics* / Q000378:metabolism; D012441:Saccharomyces cerevisiae / Q000235:genetics* / Q000378:metabolism; D029701:Saccharomyces cerevisiae Proteins / Q000737:chemistry / Q000235:genetics* / Q000378:metabolism; D016415:Sequence Alignment
D009687:Nuclear Proteins; D012337:RNA, Ribosomal, 18S; C096168:RPS14A protein, S cerevisiae; C541549:RPS14B protein, S cerevisiae; D011994:Recombinant Proteins; D012269:Ribosomal Proteins; D029701:Saccharomyces cerevisiae Proteins; D000244:Adenosine Diphosphate; D000255:Adenosine Triphosphate; D000263:Adenylate Kinase; C406456:FAP7 protein, S cerevisiae; D043583:Nucleoside-Triphosphatase