Pierre Lindenbaum Dr Pierre Lindenbaum Pierrot fr en

Pierre Lindenbaum PhD

Position
Bioinformatician
Mail
plindenbaum@yahoo.fr
Address
Integragen SA
5 rue Henri Desbruères
Genopole Campus 1, Bât G8
91030 EVRY
France
Phone: +33 (0) 160 910 900
Fax: +33 (0) 160 777 910
WWW
http://www.integragen.com
Home Page
http://www.urbigene.com
Blog
http://plindenbaum.blogspot.com
Connotea
http://www.connotea.org/user/lindenb
Other
LinkedIn
]]> Elisabeth Bouhadana Elisabeth Bouhadana Babeth Fabienne Messageot Forlot Emmanuelle Orsay University 1994 Integragen S.A. IntegraGen is a pioneer in the field of Personalized Medicine developing and delivering predictive diagnostics for a range of complex diseases. Through the discovery and validation of genetic markers using its unique genomic analysis technology (GenomeHIP) the Company is providing validated targets to the pharma and biotech industry for therapeutic development and is developing IntegraTest™ for use with patients. These tests will provide rapid and better diagnosis of disease and will help direct the choice of treatment to improve patient health outcomes.
5, rue Henri Debruere Genopole Campus 1, Bât G8
91030 EVRY
France
Phone: +33 (0) 160 910 900
Fax: +33 (0) 160 777 910
]]> Virologie Moléculaire et Structurale, Unité Mixte de Recherche, CNRS-INRA, 91198 Gif-sur-Yvette, France. Service Hospitalier Frédéric Joliot, Département de Recherche Médicale, CEA/DSV, 4 place du Général Leclerc, F-91401 Orsay, France. IDM (Immuno-Designed Molecules) Research Laboratory, University of Pierre et Marie Curie, Paris, France. Genoscope-Centre National de Séquençage and CNRS UMR-8030, 91000 Evry, France. Poncet Didier Dr Poncet Didier Rotaviruses are the major cause of viral gastroenteritis worldwide. It has been estimated that, in developing nations, over 870,000 children under the age of five die each year due to rotavirus infections. In these countries, the most severe cases are associated with malnutrition and other infectious diseases. In developed nations, rotavirus infections are rarely deadly. However, in the USA, it has been estimated that rotavirus infections result in over 50,000 hospitalizations, and result in 20 to 40 childhood deaths, annually. In addition, costs from rotavirus infections are calculated to exceed US$300 million each year in medical care alone.
The rotavirus genome is comprised of eleven molecules of double stranded RNA, each ranging in size from 3.3 to 0.6 kb. It is thought that each viral particle contains one complete set of the eleven genes. Thus, it is likely that a precise mechanism is required to control the encapsidation of each of the eleven genes.
Our laboratory is investigating different aspects of rotavirus biology with the goal of developing new vaccination strategies and antiviral approaches. Our interests include :
  • viral entry: the role of trypsin and calcium, and the structure of viral proteins.
  • viral mRNA translation: the role of the viral protein NSP3 and of cellular proteins interacting with NSP3.
  • virus assembly: the structure of viral proteins and viral complexes and their functions in viral RNA packaging.
      • ]]> Grosclaude J Dr Grosclaude Péry P Dr Péry Labbé M Dr Labbé Laude Hubert Dr Laude Hubert Petit Stéphanie Dr Petit Stéphanie Delmas Bernard Dr Delmas Bernard Bremont M Dr Bremont Béarzotti Monique Dr Béarzotti Monique Biacchesi Stéphane Dr Biacchesi Stéphane Villoing Stéphane Dr Villoing Stéphane Benmansour Abdenour Dr Benmansour Abdenour Riffault S Dr Riffault La Bonnardière C Dr La Bonnardière La Bonnardiere C Dr La Bonnardiere LeFèvre F Dr LeFèvre Lefevre F Dr Lefevre Carrat C Dr Carrat Baudoux P Dr Baudoux Charpilienne A Dr Charpilienne Aponte C Dr Aponte Gajardo R Dr Gajardo Estes M K Dr Estes M K Petitpas I Dr Petitpas Guerin-Danan Corinne Dr Guerin-Danan Nejmeddine M Dr Nejmeddine Lopez S Dr Lopez Charley Bernard Dr Charley Bernard Lefèvre François Dr Lefèvre François Boudinot Pierre Dr Boudinot Pierre Bearzotti M Dr Bearzotti Lefrère François Dr Lefrère François Le Dur Annick Dr Le Dur Annick Chapuis J Dr Chapuis Archer Fabienne Dr Archer Fabienne Moudjou Mohammed Dr Moudjou Mohammed Vilette Didier Dr Vilette Didier Delaunay T Dr Delaunay Arias C F Dr Arias C F López S Dr López Duarte M Dr Duarte Piron M Dr Piron Castagné N Dr Castagné Cohen Jean Dr Cohen Jean

      Jean Cohen (1941-2004)

      After studying physics and biochemistry and gaining a PhD degree at the University of Orsay/Paris, Jean entered the Institut National de la Recherche Agronomique in 1969, where he worked for 35 years as a virologist. Jean devoted almost his entire professional career to the study of the structure and molecular biology of rotaviruses, a major cause of infantile gastroenteritis in humans and of diarrhea in the young of a large variety of animals. Jean is rightly regarded by many as the father of the molecular biology of rotaviruses. He published/co-published the results of his work in over 100 scientific papers. His achievements were many: the discovery of the transcriptase activity of rotavirus subviral particles in vitro, description of the rotavirus genome from the early days of cloning and sequencing onwards, studies of molecular mechanisms of viral replication including functions of virus-coded nonstructural proteins and contribution to the elucidation of the atomic structure of VP6. He explored the virus-like particles produced by recombinant baculoviruses in various ways, and many of his later studies related to immune responses to rotavirus infections as well as to vaccine research. Jean was a researcher with immense insight into his subject, lots of original and creative ideas, an always searching and questioning mind and exceptional skills at the bench. He was enthusiastic for his work and stimulated many who collaborated with him. As a scientist, thinker, author, mentor, friend and colleague, he was generous, thoughtful, modest, and an inspiration to those who knew him. He listened carefully, engaged ideas, loved hardy debates, and his advice and collaboration were often sought. He was widely read and possessed a deep sense of humour. He is sorely missed by his family, friends and colleagues, but will live on in our memory.
      A. Charpilienne, U. Desselberger, M. K. Estes, D. Poncet ]]>             Berois Mabel Dr Berois Mabel Becker Michelle M Dr Becker Michelle M Vende Patrice Dr Vende Patrice Brémont M Dr Brémont Patton John T Dr Patton John T Vitour Damien Dr Vitour Damien Lavialle Françoise Dr Lavialle Françoise Chanat Eric Dr Chanat Eric Boisgard Raphaël Dr Boisgard Raphaël Evaluation of drug penetration into the brain: a double study by in vivo imaging with positron emission tomography and using an in vitro model of the human blood-brain barrier. The blood-brain barrier (BBB) permeabilities of 11 compounds were measured both in vitro with a newly developed coculture-based model of human BBB and in vivo with positron emission tomography (PET). The 11 compounds were fluoropyridinyl derivatives labeled with the positron-emitter fluorine-18, [(18)F]F-A-85380 [2-[(18)F]fluoro-3-[2(S)-2 azetidinylmethoxy]pyridine], and 10 selected N-substituted-azetidinyl and pyrrolidinyl closely related [(18)F]fluoropyridinyl derivatives (including [N"-aromatic/aliphatic]-thioureas, -ureas, and -amides). The in vitro BBB model, a new coculture system of primary human brain endothelial cells and astrocytes, was used to measure the permeability coefficient for each compound. Dynamic PET studies were performed in rats with the same compounds, and a two-compartment model analysis was used to calculate their in vivo permeability coefficients. The 11 derivatives differed in their degree of BBB passage and transport mechanism. The analysis of PET data showed a significant cerebral uptake for six derivatives, for which the in vitro evaluation indicated active influx or free diffusion. Five derivatives displayed low in vivo cerebral uptake, in agreement with the observation of an in vitro active efflux. Overall, there was a remarkable correlation between the in vitro and in vivo permeability coefficients (r = 0.99). This double study proves a close correlationship between the assessment of the BBB passage in vitro and in vivo. The in vitro model of human BBB offers the possibility of subtle discrimination of various BBB permeability degrees and transport mechanisms. Conversely, small animal PET imaging appears suitable to screen directly in vivo brain targeting of drugs or radiopharmaceutical candidates. Arrieumerlou Cécile Dr Arrieumerlou Cécile A local coupling model and compass parameter for eukaryotic chemotaxis. Chemotaxis is a cellular sensing mechanism that guides immune cells to sites of infection and leads fibroblasts to sites of injury. Here, we show in migrating primary dendritic cells and fibroblasts that the leading edge is not a uniform signaling entity, but instead consists of independent coupling units in which transient activation of PI3-kinase links to local lamellipod extension and small discrete turns in the direction of migration. These findings led to a model in which global cell polarization is independent from the chemotaxis mechanism. In this model, chemotaxis does not require spatial integration but is instead a stochastic process in which each receptor binding event within the leading edge triggers a local lamellipod extension and a small turn in the direction of migration. We show that this model and a derived "compass parameter" are sufficient to simulate the observed random migration, biased random walk, and persistent chemotactic behaviors of eukaryotic cells Bercovici Nadège Dr Bercovici Nadège Vaccination of melanoma patients using dendritic cells loaded with an allogeneic tumor cell lysate. The aim of the present phase I/II study was to evaluate the safety, immune responses and clinical activity of a vaccine based on autologous dendritic cells (DC) loaded with an allogeneic tumor cell lysate in advanced melanoma patients. DC derived from monocytes were generated in serum-free medium containing GM-CSF and IL-13 according to Good Manufacturing Practices. Fifteen patients with metastatic melanoma (stage III or IV) received four subcutaneous, intradermal, and intranodal vaccinations of both DC loaded with tumor cell lysate and DC loaded with hepatitis B surface protein (HBs) and/or tetanus toxoid (TT). No grade 3 or 4 adverse events related to the vaccination were observed. Enhanced immunity to the allogeneic tumor cell lysate and to TAA-derived peptides were documented, as well as immune responses to HBs/TT antigens. Four out of nine patients who received the full treatment survived for more than 20 months. Two patients showed signs of clinical response and received 3 additional doses of vaccine: one patient showed regression of in-transit metastases leading to complete remission. Eighteen months later, the patient was still free of disease. The second patient experienced stabilization of lung metastases for approximately 10 months. Overall, our results show that vaccination with DC loaded with an allogeneic melanoma cell lysate was feasible in large-scale and well-tolerated in this group of advanced melanoma patients. Immune responses to tumor-related antigens documented in some treated patients support further investigations to optimize the vaccine formulation. Abastado Jean-Pierre Dr Abastado Jean-Pierre Tores F Dr Tores Dr Nathalie VIONNET, MD, PhD, training in human genetics and endocrinology. Researcher at the INSERM with major focus on the genetics of type 2 diabetes and vascular complications of diabetes. Made significant contributions to the genetic dissection of type 2 diabetes with ~40 international publications in the field. Coordinator of a large Framework 5 integrated project (genetics of diabetic complications) of the European Union. Vionnet N Dr Vionnet Philippe Gesnouin Philippe Gesnouin
      Vice-President of Operations
      Mr. Gesnouin joined Integragen as Founder and Chief Information Officer in 2001. Prior to that, he was Director of Information Technology at the Xeotron Corporation (Houston, TX) and Lexicon Genetics (Houston, TX) where he supervised and developed these companies information system, data management and bio-informatics.
      Mr Gesnouin has been a research engineer and system administrator at Infobiogen, where he was instrumental in the implementation and development of France"s first centralized bioinformatics site, at the Centre for Human Polymorphic Studies, and at Genethon, where he co-developed the automation systems for data analysis for the development of the first physical and genetic maps of the human genome.
      Mr. Gesnouin obtained a Professional Master"s degree in applied computer science for biology from the University of Paris, VI, and a Research Master"s degree in animal physiology and biochemistry from the University of Paris, XI. ]]>             Philippi A Dr Philippi Ms. Philippi joined Integragen as Founder and Director of Bio-statistics and Genetic Epidemiology in 2001. Ms. Philippi has enjoyed over ten (10) years of scientific programming success working within academic and public agencies. Most recently, she held a research assistantship at the Baylor College of Medicine. As an engineer at the Center for Human Polymorphic Studies (1992-1996), and then at INSERM in Paris (1996-1998), she gained expertise in applying bio-statistical analysis to the genetics of multi-factorial disease, focusing on obesity and diabetes.Ms. Philippi has developed a series of sophisticated bio-analysis tools including a variant of the "Maximum Likelihood Binomial" method to elucidate correlative polymorphisms. Ms. Philippi obtained a Master"s in bio-informatics from the University of Paris, VI, and a Master’s in biomedical engineering from the University of Paris, XI. ]]> Hager J Dr Hager Jörg Hager, PhD
      Dr Hager has over thirty (30) publications and five (5) patents, most of which are associated with the genetics of metabolic disorders, and Integragen"s GenomeHIP technology.
      Dr Hager received his PhD from the University of Bonn in Germany. Dr Hager is the recipient of several honors and awards including an EMBO fellowship and the Young Scientist Award from the European Society for Human Genetics. ]]>             Jan Mous Dr Jan Mous, PhD
      President and Chief Executive Officer (CEO)
      Dr Mous recently joined Integragen as President and Chief Executive Officer in July 2003. Dr Mous is also a Member of the Scientific Advisory Board of Proteosys, a biotechnology company; and the Scientific Expert Committee of "First Ventury", a venture capital firm.
      Dr Mous has nearly twenty (20) years working in the bio-pharmaceuticals industry primarily at F. Hoffman-La Roche and most recently LION Bioscience.
      Dr. Mous was Global Head of Genomics Technologies (1998-2000), deputy Head CNS research (1997-1998), Head Gene Technology/Genomics (1994-1997), Head Molecular Biology (1991-1994) and Head AIDS research and Molecular Virology (1985-1994) at F.Hoffmann-La Roche Ltd, Basel, Switzerland. Also at Roche, Dr Mous initiated and managed alliances with Incyte and Affymetrix, initiated the first pharmaco- and toxico-genomics programs, and led the creation of the proteomics project.
      Dr Mous then served as Chief Scientific Officer at LION Biosciences, enjoying the firm winning the Most Successful IPO of the Year award in 2000.
      Dr Mous is author of over 80 publications in international journals. He served as a lecturer, tutor and supervisor to various universities including those of Basel, Bern, Brussels, Leuven, and Zurich. He is also a "Privatdozent" at the University of Basel. Dr Mous obtained his PhD in biochemistry from the University of Leuven (Belgium) and his "venia docendi" also in biochemistry from the University of Basel.]]>             Foglio M Dr Foglio studied molecular biology at University of California, San Diego, and he collaborated in the study of ADH expression patterns and retinol metabolism at the Burnham Institute. His work involved gene cloning, gene knockouts, and molecular modelling. He latter earned a degree in software engineering. Currently he is working on the development of a genomic database management system that facilitates the production management for SNP discovery and the implementation of customized workflows for large-scale genomic analysis. ]]> Lathrop Mark Dr Lathrop Mark Margetic Nino Dr Margetic Nino Takahashi Masazumi Dr Takahashi Masazumi I had worked 5 years on molecular immunology in Kyoto and moved to F. R. Blattner"s laboratory. After I came back to Kyoto (IIAS) I started working on the acetylcholine receptor. Since I joined CNG, I’ve been spending much time developing an automated SNP discovery tool. ]]> Matsuda Fumihiko Dr Matsuda Fumihiko Gyapay G Dr Gyapay Head of the mapping department at the French National Sequencing Center, Genoscope, in Paris where he is responsible for the human chromosome 14 sequencing, part of the Human Genome Project. Prior to joining Genoscope, Dr Gyapay has been instrumental in the first mapping of the human genome as part of the Genethon genetic mapping project. ]]> Anthouard Véronique Dr Anthouard Véronique Saurin William Dr Saurin William Petit Jean-Louis Dr Petit Jean-Louis Scarpelli Claude Dr Scarpelli Claude Weissenbach J Dr Weissenbach Lathrop G M Dr Lathrop G M Caloustian C Dr Caloustian Escary J L Dr Escary J L Gut I G Dr Gut I G Boussicault Francis Dr Boussicault Francis Tost Jörg Dr Tost Jörg Lechner D Dr Lechner Besse Celine Sauer S Dr Sauer Rousseau Francis Dr Rousseau Francis Dr. Rousseau joined Integragen as Director of Applied Research in 2002. Dr. Rousseau has worked in both small entrepreneurial biotechnology as well as large pharmaceutical firms. Most recently as Director of Genotyping at GenOdyssee, he led the construction of the ISO 9000 high-throughput SNP genotyping facility. Dr. Rousseau also worked as a researcher at Aventis Pharma, where he constructed and managed a team investigating the genetics of complex traits such as schizophrenia, Bi-Polar and Parkinson"s diseases He has also worked at ORIS-Industrie, and INSERM U393, directed by Professor Arnold Munich. Dr. Rousseau received his PhD (Honorable Mention) and Masters (Honorable Mention) in Human Genetics from the University of Pierre and Marie Curie. ]]> Demange N Dr Demange Brooks P Dr Brooks Program Director, IntegraChips
      Dr Brooks joined Integragen as founder and Director of Technology Development in 2001. Previously, Dr Brooks was a CNRS scientist at Genethon and at France"s National Sequencing Center, where he led a team to develop protein-dependent protocols for detecting DNA sequence variations, as well as performing in vitro studies on DNA repair. At the Institut Jacques Monod in Paris, he directed the biochemistry group in the Laboratory of Mutagenesis and was among the first in the field of DNA mismatch repair to establish a system for studying mismatch repair in a model organism. Dr Brooks obtained his Ph.D. in biochemistry at the University of California at Berkeley under the direction of Stuart Linn and Bruce Ames. ]]>             Abdel Benajou

      Abdel Benajou

      Works at Integragen]]>             Alain Astgen

      Alain Astgen

      Worked at CNG]]>             Bruno Copin

      Bruno Copin

      Works at Integragen]]>             Celine Vazart

      Celine Vazart

      Works at Integragen]]>             Charles Marcaillou

      Charles Marcaillou

      Works at Integragen]]>             Claudia Sannka

      Claudia Sannka

      Works at Integragen]]>             Danielle Bassez

      Danielle Bassez

      Works at Integragen]]>             Dave Brady David brings over 12 years of experience in successful deal making and alliance management. Prior to joining IntegraGen, David was Associate Director for International Corporate Development at Biogen Idec, based in their UK offices, where he steered business development opportunities for Biogen within Europe and led the resulting due diligence and negotiations. He has experience of leading and closing complex negotiations at a range of levels, from pre-clinical products to technology platforms and late-stage products. Before working at Biogen Idec, David was Commercial Manager for Lonza Biologics plc., part of the Alusuisse Lonza group providing contract management services to the biopharmaceutical industry, and Contracts Manager at Celltech Biologics Inc. in New Hampshire, USA. He moved into the Sales and Marketing Team at Celltech from a role as a development scientist. He has a BSc in Biochemistry from the University of Liverpool and an MBA from Warwick Business School. ]]> Ekrame Ayari

      Ekrame Ayari

      Works at Integragen]]>             Jean-Paul Saraiva

      Jean-Paul Saraiva

      Works at Integragen]]>             Jerome Carayol

      Jerome Carayol

      Works at Integragen]]>             Jolanta Luberda

      Jolanta Luberda

      Works at Integragen]]>             Karine Fontaine

      Karine Fontaine

      Works at Integragen]]>             Livia Rival

      Livia Rival

      Works at Integragen]]>             Maud Vanpeene

      Maud Vanpeene

      Works at Integragen]]>             Patrick Court

      Patrick Court

      Works at Integragen]]>             Paul Guermonprez

      Paul Guermonprez

      Works at Integragen]]>             Stephane Roy

      Stephane Roy

      Works at Integragen]]>             Stephanie Maillard

      Stephanie Maillard

      Works at Integragen]]>             Virginie Decaulne

      Virginie Decaulne

      Works at Integragen]]>             Haplotypes in the gene encoding protein kinase c-beta (PRKCB1) on chromosome 16 are associated with autism.Mol Psychiatry 10 (10): 2005 Oct p. 950-60

      A Philippi, E Roschmann, F Tores, P Lindenbaum, A Benajou, L Germain-Leclerc, C Marcaillou, K Fontaine, M Vanpeene, S Roy, S Maillard, V Decaulne, J P Saraiva, P Brooks, F Rousseau and J Hager

      Haplotypes in the gene encoding protein kinase c-beta (PRKCB1) on chromosome 16 are associated with autism.

      PMID:16027742

      Autism is a developmental disorder characterized by impairments in social interaction and communication associated with repetitive patterns of interest or behavior. Autism is highly influenced by genetic factors. Genome-wide linkage and candidate gene association approaches have been used to try and identify autism genes. A few loci have repeatedly been reported linked to autism. Several groups reported evidence for linkage to a region on chromosome 16p. We have applied a direct physical identity-by-descent (IBD) mapping approach to perform a high-density (0.85 megabases) genome-wide linkage scan in 116 families from the AGRE collection. Our results confirm linkage to a region on chromosome 16p with autism. High-resolution single-nucleotide polymorphism (SNP) genotyping and analysis of this region show that haplotypes in the protein kinase c-beta gene are strongly associated with autism. An independent replication of the association in a second set of 167 trio families with autism confirmed our initial findings. Overall, our data provide evidence that the PRKCB1 gene on chromosome 16p may be involved in the etiology of autism.

      ]]>       RoXaN, a novel cellular protein containing TPR, LD, and zinc finger motifs, forms a ternary complex with eukaryotic initiation factor 4G and rotavirus NSP3. J Virol 78 (8):2004 Apr p. 3851-62

      Damien Vitour, Pierre Lindenbaum, Patrice Vende, Michelle M Becker and Didier Poncet

      RoXaN, a novel cellular protein containing TPR, LD, and zinc finger motifs, forms a ternary complex with eukaryotic initiation factor 4G and rotavirus NSP3.

      PMID:15047801

      Rotavirus mRNAs are capped but not polyadenylated, and viral proteins are translated by the cellular translation machinery. This is accomplished through the action of the viral nonstructural protein NSP3, which specifically binds the 3' consensus sequence of viral mRNAs and interacts with the eukaryotic translation initiation factor eIF4G I. To further our understanding of the role of NSP3 in rotavirus replication, we looked for other cellular proteins capable of interacting with this viral protein. Using the yeast two-hybrid assay, we identified a novel cellular protein-binding partner for rotavirus NSP3. This 110-kDa cellular protein, named RoXaN (rotavirus X protein associated with NSP3), contains a minimum of three regions predicted to be involved in protein-protein or nucleic acid-protein interactions. A tetratricopeptide repeat region, a protein-protein interaction domain most often found in multiprotein complexes, is present in the amino-terminal region. In the carboxy terminus, at least five zinc finger motifs are observed, further suggesting the capacity of RoXaN to bind other proteins or nucleic acids. Between these two regions exists a paxillin leucine-aspartate repeat (LD) motif which is involved in protein-protein interactions. RoXaN is capable of interacting with NSP3 in vivo and during rotavirus infection. Domains of interaction were mapped and correspond to the dimerization domain of NSP3 (amino acids 163 to 237) and the LD domain of RoXaN (amino acids 244 to 341). The interaction between NSP3 and RoXaN does not impair the interaction between NSP3 and eIF4G I, and a ternary complex made of NSP3, RoXaN, and eIF4G I can be detected in rotavirus-infected cells, implicating RoXaN in translation regulation.

      ]]>       CloneIt: finding cloning strategies, in-frame deletions and frameshifts. Bioinformatics 14 (5): 1998 Jun p. 465-6

      P Lindenbaum

      CloneIt: finding cloning strategies, in-frame deletions and frameshifts.

      PMID:9682060

      MOTIVATION: The CloneIt program searches for sub-cloning strategies, in-frame deletions and frameshifts within a plasmid sequence. AVAILABILITY: The program, written in ANSI-C language, is available at http://locus.jouy.inra.fr/soft/cloneit/clonei t.html CONTACT: lindenb@biotec.jouy.inra.fr

      ]]>       In vivo and in vitro phosphorylation of rotavirus NSP5 correlates with its localization in viroplasms. J Virol 71 (1):1997 Jan p. 34-41

      D Poncet, P Lindenbaum, R L'Haridon and J Cohen

      In vivo and in vitro phosphorylation of rotavirus NSP5 correlates with its localization in viroplasms.

      PMID:8985320

      NSP5 (NS26), the product of rotavirus gene 11, is a phosphoprotein whose role in the virus replication cycle is unknown. To gain further insight into its function, we obtained monoclonal antibodies against the baculovirus-expressed protein. By immunoprecipitation and immunoblotting experiments, we showed that (i) NSP5 appears in many different phosphorylated forms in rotavirus-infected cells; (ii) immunoprecipitated NSP5 from rotavirus-infected cells can be phosphorylated in vitro by incubation with ATP; (iii) NSP5, produced either by transient transfection of rotavirus gene 11 or by infection by gene 11 recombinant vaccinia virus or baculovirus, can be phosphorylated in vivo and in vitro; (iv) NSP5 expressed in Escherichia coli is phosphorylated in vitro, and thus NSP5 is a potential protein kinase; and (v) NSP5 forms dimers and interacts with NSP2. The intracellular localization of NSP5 in the course of rotavirus infection and after transient expression in COS7 cells has also been investigated. In rotavirus-infected cells, NSP5 is localized in viroplasms, but it is widespread throughout the cytoplasm of transfected COS7 cells. NSP5 produced by transfected COS7 cells did not acquire the multiphosphorylated forms observed in rotavirus-infected COS7 cells. Thus, there is a tight correlation between the localization of NSP5 in the viroplasms and its protein kinase activity in vivo or in vitro. Our results suggest that cellular or viral cofactors are indispensable to fully phosphorylate NSP5 and to reach its intracellular localization.

      ]]>