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Journal of Virology, December 2002, p. 12866-12876, Vol. 76, No. 24
0022-538X/02/$04.00+0     DOI: 10.1128/JVI.76.24.12866-12876.2002
Copyright © 2002, American Society for Microbiology. All Rights Reserved.

Endocytosis Is a Critical Step in Entry of Subgroup B Avian Leukosis Viruses

Felipe Diaz-Griffero, Steven Ari Hoschander, and Jürgen Brojatsch*

Department of Microbiology and Immunology, Albert Einstein College of Medicine, Bronx, New York 10461

Received 13 May 2002/ Accepted 13 September 2002

The avian leukosis virus (ALV) entry mechanism is controversial, with evidence for and against a low-pH requirement for viral fusion. To further address this question, we tested the entry of human immunodeficiency virus type 1 (HIV-1) pseudotyped with the envelope protein of subgroup B ALV (ALV-B) in the presence of three different lysosomotropic agents. These lysosomotropic agents were able to block the entry of wild-type and pseudotyped ALV-B in two different cell lines, strongly suggesting that ALV-B requires a low-pH step for entry. ALV-B and pH-dependent Semliki Forest virus (SFV) entered cells with slower uptake kinetics than HIV-1, which is pH independent. These slow uptake rates support the theory that ALV-B utilizes endocytic pathways to enter cells. Using immunofluorescence and electron microscopy analysis, we visualized the colocalization of virus particles with the endosomal marker transferrin and demonstrated virus particles in clathrin-coated vesicles and endosome-like structures. Surprisingly, a low-pH treatment did not overcome the inhibition of ALV-B entry by lysosomotropic agents. This indicates that, in contrast to SFV, ALV-B is unable to fuse at the cellular surface, even at a low pH. Taken together, our findings suggest that endocytosis and a subsequent low-pH step are critical for successful ALV-B infection.

* Corresponding author. Mailing address: Department of Microbiology and Immunology, Albert Einstein College of Medicine, 1300 Morris Park Ave., Bronx, NY 10461. Phone: (718) 430-3079. Fax: (718) 430-8711. E-mail: brojatsc{at}aecom.yu.edu.

Journal of Virology, December 2002, p. 12866-12876, Vol. 76, No. 24
0022-538X/02/$04.00+0     DOI: 10.1128/JVI.76.24.12866-12876.2002
Copyright © 2002, American Society for Microbiology. All Rights Reserved.



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