| HOME | HELP | FEEDBACK | SUBSCRIPTIONS | ARCHIVE | SEARCH | TABLE OF CONTENTS |
|
|
|
|||||||
|
|||||||||
International Centre for Genetic Engineering and Biotechnology, Area Science Park, Padriciano 99, 34012-Trieste, Italy
Author for correspondence: Oscar Burrone. Fax + 39 40 226555, e-mail burrone{at}icgeb.trieste.it
NSP5 (non-structural protein 5) is one of two proteins encoded by genome segment 11 of group A rotaviruses. In virus-infected cells NSP5 accumulates in the virosomes and is found as two polypeptides with molecular masses of 26 and 28 kDa (26K and 28K proteins). NSP5 has been previously shown to be post-translationally modified by the addition of O-linked monosaccharide residues of N-acetylglucosamine and also by phosphorylation. We have now found that, as a consequence of phosphorylation, a complex modification process gives rise to previously unidentified forms of NSP5, with molecular masses of up to 34 kDa. Treatment with phosphatases of NSP5 obtained from virus-infected cells produced a single band of 26 kDa. NSP5 could be phosphorylated in vitro by incubation of immuno-precipitates with [
-32P]ATP, producing mainly phosphorylated products of 28 and 32–34 kDa (32–34K). In both in vivo and in vitro phosphorylated NSP5, phosphates were only found attached via serine and threonine residues. The in vitro translated NSP5 precursor polypeptide, molecular mass 25 kDa (25K), could also be phosphorylated and transformed into a 28K protein by incubation with extracts obtained from virus-infected cells, but not from non-infected cells. In addition, NSP5 labelled in vivo with [1,6-3H]glucosamine showed mainly the presence of the 26K and 28K proteins (converted to 26K by protein phosphatase treatment) suggesting that the type of protein produced is regulated according to the level of phosphorylation and/or O-glycosylation. The results also suggest that NSP5 is autophosphorylated.
Received 11 March 1996;
accepted 24 May 1996.
This article has been cited by other articles:
|
|
 |
|
  D. Martin, M. Duarte, J. Lepault, and D. Poncet Sequestration of Free Tubulin Molecules by the Viral Protein NSP2 Induces Microtubule Depolymerization during Rotavirus Infection J. Virol., March 1, 2010; 84(5): 2522 - 2532. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
M. Kumar, H. Jayaram, R. Vasquez-Del Carpio, X. Jiang, Z. F. Taraporewala, R. H. Jacobson, J. T. Patton, and B. V. V. Prasad Crystallographic and Biochemical Analysis of Rotavirus NSP2 with Nucleotides Reveals a Nucleoside Diphosphate Kinase-Like Activity J. Virol., November 15, 2007; 81(22): 12272 - 12284. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
A. Sen, N. Sen, and E. R. Mackow The Formation of Viroplasm-Like Structures by the Rotavirus NSP5 Protein Is Calcium Regulated and Directed by a C-Terminal Helical Domain J. Virol., November 1, 2007; 81(21): 11758 - 11767. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 M. Campagna, M. Budini, F. Arnoldi, U. Desselberger, J. E. Allende, and O. R. Burrone Impaired hyperphosphorylation of rotavirus NSP5 in cells depleted of casein kinase 1{alpha} is associated with the formation of viroplasms with altered morphology and a moderate decrease in virus replication J. Gen. Virol., October 1, 2007; 88(10): 2800 - 2810. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
F. Arnoldi, M. Campagna, C. Eichwald, U. Desselberger, and O. R. Burrone Interaction of Rotavirus Polymerase VP1 with Nonstructural Protein NSP5 Is Stronger than That with NSP2 J. Virol., March 1, 2007; 81(5): 2128 - 2137. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
M. Campagna, O. R. Burrone, A. Sen, and E. R. Mackow Fusion of Tags Induces Spurious Phosphorylation of Rotavirus NSP5 J. Virol., August 15, 2006; 80(16): 8283 - 8285. [Full Text] [PDF]
|
|
|
|
|
|
A. Sen, D. Agresti, and E. R. Mackow Hyperphosphorylation of the Rotavirus NSP5 Protein Is Independent of Serine 67 or NSP2, and the Intrinsic Insolubility of NSP5 Is Regulated by Cellular Phosphatases J. Virol., February 15, 2006; 80(4): 1807 - 1816. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
J. Modrof, K. Lymperopoulos, and P. Roy Phosphorylation of Bluetongue Virus Nonstructural Protein 2 Is Essential for Formation of Viral Inclusion Bodies J. Virol., August 1, 2005; 79(15): 10023 - 10031. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
T. Lopez, M. Rojas, C. Ayala-Breton, S. Lopez, and C. F. Arias Reduced expression of the rotavirus NSP5 gene has a pleiotropic effect on virus replication J. Gen. Virol., June 1, 2005; 86(6): 1609 - 1617. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
M. Campagna, C. Eichwald, F. Vascotto, and O. R. Burrone RNA interference of rotavirus segment 11 mRNA reveals the essential role of NSP5 in the virus replicative cycle J. Gen. Virol., May 1, 2005; 86(5): 1481 - 1487. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 C. Eichwald, G. Jacob, B. Muszynski, J. E. Allende, and O. R. Burrone Uncoupling substrate and activation functions of rotavirus NSP5: Phosphorylation of Ser-67 by casein kinase 1 is essential for hyperphosphorylation PNAS, November 16, 2004; 101(46): 16304 - 16309. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
F. Vascotto, M. Campagna, M. Visintin, A. Cattaneo, and O. R. Burrone Effects of intrabodies specific for rotavirus NSP5 during the virus replicative cycle J. Gen. Virol., November 1, 2004; 85(11): 3285 - 3290. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 R. V.-D. Carpio, F. D. Gonzalez-Nilo, H. Jayaram, E. Spencer, B. V. V. Prasad, J. T. Patton, and Z. F. Taraporewala Role of the Histidine Triad-like Motif in Nucleotide Hydrolysis by the Rotavirus RNA-packaging Protein NSP2 J. Biol. Chem., March 12, 2004; 279(11): 10624 - 10633. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
C. Eichwald, J. F. Rodriguez, and O. R. Burrone Characterization of rotavirus NSP2/NSP5 interactions and the dynamics of viroplasm formation J. Gen. Virol., March 1, 2004; 85(3): 625 - 634. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
K. V. K. Mohan, J. Muller, and C. D. Atreya The N- and C-Terminal Regions of Rotavirus NSP5 Are the Critical Determinants for the Formation of Viroplasm-Like Structures Independent of NSP2 J. Virol., November 15, 2003; 77(22): 12184 - 12192. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
M. Berois, C. Sapin, I. Erk, D. Poncet, and J. Cohen Rotavirus Nonstructural Protein NSP5 Interacts with Major Core Protein VP2 J. Virol., February 1, 2003; 77(3): 1757 - 1763. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
P. Vende, Z. F. Taraporewala, and J. T. Patton RNA-Binding Activity of the Rotavirus Phosphoprotein NSP5 Includes Affinity for Double-Stranded RNA J. Virol., May 15, 2002; 76(10): 5291 - 5299. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
C. Eichwald, F. Vascotto, E. Fabbretti, and O. R. Burrone Rotavirus NSP5: Mapping Phosphorylation Sites and Kinase Activation and Viroplasm Localization Domains J. Virol., April 1, 2002; 76(7): 3461 - 3470. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 J. A. HANOVER Glycan-dependent signaling: O-linked N-acetylglucosamine FASEB J, September 1, 2001; 15(11): 1865 - 1876. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
M. A. Torres-Vega, R. A. Gonzalez, M. Duarte, D. Poncet, S. Lopez, and C. F. Arias The C-terminal domain of rotavirus NSP5 is essential for its multimerization, hyperphosphorylation and interaction with NSP6 J. Gen. Virol., March 1, 2000; 81(3): 821 - 830. [Abstract] [Full Text]
|
|
|
|
|
|
S. Shen, T. A. McKee, Z. D. Wang, U. Desselberger, and D. X. Liu Sequence analysis and in vitro expression of genes 6 and 11 of an ovine group B rotavirus isolate, KB63: evidence for a non-defective, C-terminally truncated NSP1 and a phosphorylated NSP5 J. Gen. Virol., August 1, 1999; 80(8): 2077 - 2085. [Abstract] [Full Text]
|
|
|
|
|
|
E Fabbretti, I Afrikanova, F Vascotto, and O. Burrone Two non-structural rotavirus proteins, NSP2 and NSP5, form viroplasm- like structures in vivo J. Gen. Virol., February 1, 1999; 80(2): 333 - 339. [Abstract]
|
|
|
|
|
|
J. Blackhall, M. Munoz, A. Fuentes, and G. Magnusson Analysis of Rotavirus Nonstructural Protein NSP5 Phosphorylation J. Virol., August 1, 1998; 72(8): 6398 - 6405. [Abstract] [Full Text] [PDF]
|
|
| HOME | HELP | FEEDBACK | SUBSCRIPTIONS | ARCHIVE | SEARCH | TABLE OF CONTENTS |
| INT J SYST EVOL MICROBIOL | MICROBIOLOGY | J GEN VIROL |
| J MED MICROBIOL | ALL SGM JOURNALS | |
