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Journal of Virology, December 2004, p. 13627-13636, Vol. 78, No. 24
0022-538X/04/$08.00+0     DOI: 10.1128/JVI.78.24.13627-13636.2004
Copyright © 2004, American Society for Microbiology. All Rights Reserved.

C-Terminal nsP1a Protein of Human Astrovirus Colocalizes with the Endoplasmic Reticulum and Viral RNA

Susana Guix, Santiago Caballero, Albert Bosch,* and Rosa M. Pintó

Enteric Virus Laboratory, Department of Microbiology, University of Barcelona, Barcelona, Spain

Received 12 March 2004/ Accepted 5 August 2004

Computational and biological approaches were undertaken to characterize the role of the human astrovirus nonstructural protein nsP1a/4, located at the C-terminal fragment of nsP1a. Computer analysis reveals sequence similarities to other nonstructural viral proteins involved in RNA replication and/or transcription and allows the identification of a glutamine- and proline-rich region, the prediction of many phosphorylation and O-glycosylation sites, and the occurrence of a KKXX-like endoplasmic reticulum retention signal. Immunoprecipitation analysis with an antibody against a synthetic peptide of the nsP1a/4 sequence detected polyprotein precursors of 160, 75, and 38 to 40 kDa as well as five smaller proteins in the range of 21 to 27 kDa. Immunofluorescence labeling showed that the nsP1a/4 protein is accumulated at the perinuclear region, in association with the endoplasmic reticulum and the viral RNA. These results suggest the involvement of nsP1a/4 protein in the RNA replication process in endoplasmic reticulum-derived intracellular membranes.

* Corresponding author. Mailing address: Department of Microbiology, University of Barcelona, Avda Diagonal 645, 08028 Barcelona, Spain. Phone: (34) 934034620. Fax: (34) 934034629. E-mail: abosch{at}ub.edu.

Journal of Virology, December 2004, p. 13627-13636, Vol. 78, No. 24
0022-538X/04/$08.00+0     DOI: 10.1128/JVI.78.24.13627-13636.2004
Copyright © 2004, American Society for Microbiology. All Rights Reserved.



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