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Journal of Virology, April 2001, p. 3771-3778, Vol. 75, No. 8
0022-538X/01/$04.00+0   DOI: 10.1128/JVI.75.8.3771-3778.2001
Copyright © 2001, American Society for Microbiology. All rights reserved.

Actin Rearrangement-Inducing Factor of Baculoviruses Is Tyrosine Phosphorylated and Colocalizes to F-Actin at the Plasma Membrane

Stephan Dreschers,1 Renza Roncarati,2 and Dagmar Knebel-Mörsdorf1,*

Max Planck Institute for Neurological Research and Department of Neurology, University of Cologne, Cologne, Germany,1 and Department of Medicine and Public Health, University of Verona, Verona, Italy2

Received 20 October 2000/Accepted 8 January 2001

In previous studies we have identified actin rearrangement-inducing factor 1 as an early gene product of Autographa californica multicapsid nuclear polyhedrosis virus that is involved in the remodeling of the actin cytoskeleton. We have constructed viral recombinants with a mutated Arif-1 open reading frame that confirm the causal link of Arif-1 expression and the actin rearrangement observed as accumulation of F-actin at the plasma membrane at 3 to 7 h postinfection. Infection with Arif mutant viruses leads to the loss of actin accumulation at the plasma membrane in TN-368 cells, although in the course of infection, early actin cables and nuclear F-actin are observed as in wild-type-infected cells. By immunofluorescence studies, we have demonstrated the localization of Arif-1 at the plasma membrane, and confocal imaging reveals the colocalization to F-actin. Accordingly, the ~47-kDa Arif-1 protein is observed exclusively in membrane fractions prepared at 4 to 48 h postinfection, with a decrease at 24 h postinfection. Phosphatase treatment suggests that Arif-1 is modified by phosphorylation. Antibodies against phosphotyrosine precipitate Arif-1 from membrane fractions, indicating that Arif-1 becomes tyrosine phosphorylated during the early and late phases of infection. In summary, our results indicate that functional Arif-1 is tyrosine phosphorylated and is located at the plasma membrane as a component of the actin rearrangement-inducing complex.

* Corresponding author. Mailing address: Max Planck Institute for Neurological Research, Gleuelerstrasse 50, 50931 Cologne, Germany. Phone: 49-221-4726261. Fax: 49-221-4726298. E-mail: D.Moersdorf{at}pet.mpin-koeln.mpg.de.

Journal of Virology, April 2001, p. 3771-3778, Vol. 75, No. 8
0022-538X/01/$04.00+0   DOI: 10.1128/JVI.75.8.3771-3778.2001
Copyright © 2001, American Society for Microbiology. All rights reserved.


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