Analysis of the Ribosomal Protein S19 Interactome

Orrú, Stefania; Aspesi, Anna; Armiraglio, Marta; Caterino, Marianna; Loreni, Fabrizio; Ruoppolo, Margherita; Santoro, Claudio; Dianzani, Irma

doi:10.1074/mcp.m600156-mcp200

Cited by 53 publications

(65 citation statements)

References 47 publications

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“…In vitro, interaction between RPS19 and PIM-1 leads to its phosphorylation [Chiocchetti et al, 2005]. The entire RPS19 interactome has been published: interestingly, most interactors are nucleolar, and include many RPs, preribosome components, and helicases [Orrù et al, 2007]. This is consistent with the finding that RPS19 is essential for the maturation of 18S rRNA [Flygare et al, 2007;Choesmel et al, 2007;Idol et al, 2007] and assembly of pre-40S particles [Léger-Silvestre et al, 2005].…”

Section: Rps19supporting

confidence: 77%

RPS19 mutations in patients with Diamond-Blackfan anemia

et al. 2008

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Communicated by Maria Rita Passos-BuenoDiamond-Blackfan anemia (DBA) is an inherited disease characterized by pure erythroid aplasia. Thirty percent (30%) of patients display malformations, especially of the hands, face, heart, and urogenital tract. DBA has an autosomal dominant pattern of inheritance. De novo mutations are common and familial cases display wide clinical heterogeneity. Twenty-five percent (25%) of patients carry a mutation in the ribosomal protein (RP) S19 gene, whereas mutations in RPS24, RPS17, RPL35A, RPL11, and RPL5 are rare. These genes encode for structural proteins of the ribosome. A link between ribosomal functions and erythroid aplasia is apparent in DBA, but its etiology is not clear. Most authors agree that a defect in protein synthesis in a rapidly proliferating tissue, such as the erythroid bone marrow, may explain the defective erythropoiesis. A total of 77 RPS19 mutations have been described. Most are whole gene deletions, translocations, or truncating mutations (nonsense or frameshift), suggesting that haploinsufficiency is the basis of DBA pathology. A total of 22 missense mutations have also been described and several works have provided in vitro functional data for the mutant proteins. This review looks at the data on all these mutations, proposes a functional classification, and describes six new mutations. It is shown that patients with RPS19 mutations display a poorer response to steroids and a worse long-term prognosis compared to other DBA patients. Hum Mutat 29(7), [911][912][913][914][915][916][917][918][919][920] 2008.

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Section: Rps19supporting

confidence: 77%

RPS19 mutations in patients with Diamond-Blackfan anemia

et al. 2008

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“…Recently RPS19 has been found to interact with cellular protein from diverse protein families, including NTPases, hydrolases, isomerases, kinases, splicing factors, transcription factors, transferases, and DNA/ RNA-binding proteins (64). Although the biological significance of such diverse interactions of RPS19 is still unknown, it is likely that the N-RPS19 interaction will have an effect upon other functions of RPS19.…”

Section: Discussionmentioning

confidence: 99%

Characterization of the Interaction between Hantavirus Nucleocapsid Protein (N) and Ribosomal Protein S19 (RPS19)

Cheng

Haque

Rimmer

et al. 2011

Journal of Biological Chemistry

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Hantaviruses, members of the Bunyaviridae family, are negative-stranded emerging RNA viruses and category A pathogens that cause serious illness when transmitted to humans through aerosolized excreta of infected rodent hosts. Hantaviruses have evolved a novel translation initiation mechanism, operated by nucleocapsid protein (N), which preferentially facilitates the translation of viral mRNAs. N binds to the ribosomal protein S19 (RPS19), a structural component of the 40 S ribosomal subunit. In addition, N also binds to both the viral mRNA 5 cap and a highly conserved triplet repeat sequence of the viral mRNA 5 UTR. The simultaneous binding of N at both the terminal cap and the 5 UTR favors ribosome loading on viral transcripts during translation initiation. We characterized the binding between N and RPS19 and demonstrate the role of the N-RPS19 interaction in N-mediated translation initiation mechanism. We show that N specifically binds to RPS19 with high affinity and a binding stoichiometry of 1:1. The N-RPS19 interaction is an enthalpy-driven process. RPS19 undergoes a conformational change after binding to N. Using T7 RNA polymerase, we synthesized the hantavirus S segment mRNA, which matches the transcript generated by the viral RNA-dependent RNA polymerase in cells. We show that the N-RPS19 interaction plays a critical role in the translation of this mRNA both in cells and rabbit reticulocyte lysates. Our results demonstrate that the N-mediated translation initiation mechanism, which lures the host translation machinery for the preferential translation of viral transcripts, primarily depends on the N-RPS19 interaction. We suggest that the N-RPS19 interaction is a novel target to shut down the N-mediated translation strategy and hence virus replication in cells.Hantaviruses, members of the Bunyaviridae family, are category A pathogens and causative agents of two emerging diseases: hemorrhagic fever with renal syndrome and hantavirus cardiopulmonary syndrome with mortalities of 15 and 50%, respectively (1, 2). Hantaviruses are transmitted to humans through aerosolized excreta of infected rodent hosts. The spherical hantavirus particles harbor three negative sense genomic RNA segments (S, L, and M) within a lipid bilayer (3). The mRNAs derived from S, L, and M segments encode viral nucleocapsid protein (N), 2 viral RNA-dependent RNA polymerase (RdRp), and glycoprotein precursor, respectively. The glycoprotein precursor is cleaved at a conserved WAASA site, and two glycoproteins, Gn and Gc, are generated (4). The characteristic feature of the hantaviral genome is the partially complementary sequence at the 5Ј and 3Ј termini of each of the three genome segments that undergo base pairing and form panhandle structures (5-7). N is a multifunctional protein playing vital roles in multiple processes of the virus replication cycle and enters the host cell along with viral capsid during infection. N has been found to undergo trimerization both in vivo and in vitro (8 -20). During encapsidation, N specifically recogniz...

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“…Proteins were separated on 12% sodium dodecyl sulfatepolyacrylamide gel, and transferred onto nitrocellulose Protran membrane (Schleicher and Schuell, Milan, Italy), and incubated with the following primary antibodies and antisera: mouse monoclonal antibody specific for RPS19 (Orru et al, 2007), mouse monoclonal anti-GAPDH (Chemicon, Milan, Italy), rabbit polyclonal anti-RPS6 (Cell Signaling, Boston, MA, USA), rabbit polyclonal anti-b-actin (Sigma, Milan, Italy), mouse monoclonal anti-PIM1 (Santa Cruz, Heidelberg, Germany), mouse monoclonal anti-p53 (Santa Cruz), rabbit polyclonal anti-L7a (kindly provided by Giulia Russo, Naples, Italy), rabbit polyclonal anti-AKT (Santa Cruz), mouse monoclonal anti-p27 Kip1 (BD, Milan, Italy), rabbit polyclonal anti-phospho-p27 Kip1 (threonine157; R&D Systems (Minneapolis, MN, USA), kindly provided by Giuseppe Viglietto, Catanzaro, Italy), rabbit polyclonal anti-eIF4E (Cell Signaling) and rabbit polyclonal anti-b-tubulin (Santa Cruz). Primary antibodies were revealed using horseradish peroxidase-conjugated goat anti-rabbit or anti-mouse Ab (Jackson Immunoresearch, Suffolk, UK) and the ECL chemiluminescence detection system (Pierce, Rockford, IL, USA).…”

Section: Western Blottingmentioning

confidence: 99%

PIM1 kinase is destabilized by ribosomal stress causing inhibition of cell cycle progression

et al. 2010

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PIM1 is a constitutively active serine/threonine kinase regulated by cytokines, growth factors and hormones. It has been implicated in the control of cell cycle progression and apoptosis and its overexpression has been associated with various kinds of lymphoid and hematopoietic malignancies. The activity of PIM1 is dependent on the phosphorylation of several targets involved in transcription, cell cycle and apoptosis. We have recently observed that PIM1 interacts with ribosomal protein (RP)S19 and cosediments with ribosomes. Defects in ribosome synthesis (ribosomal stress) have been shown to activate a p53-dependent growth arrest response. To investigate if PIM1 could have a role in the response to ribosomal stress, we induced ribosome synthesis alterations in TF-1 and K562 erythroid cell lines. We found that RP deficiency, induced by RNA interference or treatment with inhibitor of nucleolar functions, causes a drastic destabilization of PIM1. The lower level of PIM1 induces an increase in the cell cycle inhibitor p27 Kip1 and blocks cell proliferation even in the absence of p53. Notably, restoring PIM1 level by transfection causes a recovery of cell growth. Our data indicate that PIM1 may act as a sensor for ribosomal stress independently of or in concert with the known p53-dependent mechanisms.

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Analysis of the Ribosomal Protein S19 Interactome

Cited by 53 publications

References 47 publications

RPS19 mutations in patients with Diamond-Blackfan anemia

RPS19 mutations in patients with Diamond-Blackfan anemia

Characterization of the Interaction between Hantavirus Nucleocapsid Protein (N) and Ribosomal Protein S19 (RPS19)

PIM1 kinase is destabilized by ribosomal stress causing inhibition of cell cycle progression

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