JVI Accepts, published online ahead of print on 4 November 2009

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J. Virol. doi:10.1128/JVI.01660-09
Copyright (c) 2009, American Society for Microbiology and/or the Listed Authors/Institutions. All Rights Reserved.

The adeno-associated virus (AAV) small Rep proteins are modified with at least two types of polyubiquitination

K. David Farris, Olufemi Fasina, Loretta Sukhu, Long Li, and David J. Pintel^*

Department of Molecular Microbiology and Immunology, University of Missouri-Columbia, School of Medicine, Bond Life Sciences Center, Columbia, MO

^* To whom correspondence should be addressed. Email: pinteld{at}missouri.edu.

Adeno-associated virus (AAV) types 2 and 5 Rep52 and Rep40 were polyubiquitinated during AAV/adenovirus type 5 (Ad5) coinfection and during transient transfection either in the presence or absence of Ad5 E4orf6 and E1b-55k. Polyubiquitination of small Rep proteins via lysine 48 (K48) linkages, normally associated with targeting of proteins for proteasomal degradation, was detected only in the presence of E4orf6. The small Rep proteins were ubiquitinated via lysine 63 (K63) following transfection in either the presence or absence of E4orf6, or following coinfection with Ad5. E4orf6/E1b-55k-dependent K48-specific polyubiquitination of small Rep proteins could be inhibited using siRNA to cullin 5.