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Journal of Virology, December 2006, p. 11881-11886, Vol. 80, No. 23
0022-538X/06/$08.00+0     doi:10.1128/JVI.01471-06
Copyright © 2006, American Society for Microbiology. All Rights Reserved.

Cryoelectron Microscopy of Protein IX-Modified Adenoviruses Suggests a New Position for the C Terminus of Protein IX{triangledown}

Michael P. Marsh,1,2,{dagger} Samuel K. Campos,5,7,{dagger},{ddagger} Matthew L. Baker,2 Christopher Y. Chen,3 Wah Chiu,1,2 and Michael A. Barry3,4,5,6,8*

Program in Structural and Computational Biology and Molecular Biophysics,1 National Center for Macromolecular Imaging, Verna and Marrs McLean Department of Biochemistry and Molecular Biology,2 Department of Molecular and Human Genetics,3 Department of Immunology,4 Center for Cell and Gene Therapy, Baylor College of Medicine, Houston, Texas 77030,5 Departments of Bioengineering,6 Biochemistry and Cell Biology, Rice University, Houston, Texas,7 Department of Internal Medicine, Division of Infectious Diseases, Translational Immunovirology Program, Mayo Clinic, Rochester, Minnesota8

Received 11 July 2006/ Accepted 13 September 2006

Recombinant human adenovirus is a useful gene delivery vector for clinical gene therapy. Minor capsid protein IX of adenovirus has been of recent interest since multiple studies have shown that modifications can be made to its C terminus to alter viral tropism or add molecular tags and/or reporter proteins. We examined the structure of an engineered adenovirus displaying the enhanced green fluorescent protein (EGFP) fused to the C terminus of protein IX. Cryoelectron microscopy and reconstruction localized the C-terminal EGFP fusion between the H2 hexon and the H4 hexon, positioned between adjacent facets, directly above the density previously assigned as protein IIIa. The original assignment of IIIa was based largely on indirect evidence, and the data presented herein support the reassignment of the IIIa density as protein IX.

* Corresponding author. Mailing address: 200 First St. SW, Guggenheim 5-42A, Rochester, MN 55902. Phone: (507) 288-1688. Fax: (713) 798-5959. E-mail: barry.michael{at}mayo.edu.

{triangledown} Published ahead of print on 20 September 2006.

{dagger} These authors contributed equally to this work.

{ddagger} Present address: Department of Molecular Genetics and Microbiology, University of New Mexico School of Medicine, 915 Camino de Salud, CRF 303, Albuquerque, NM 87131.

Journal of Virology, December 2006, p. 11881-11886, Vol. 80, No. 23
0022-538X/06/$08.00+0     doi:10.1128/JVI.01471-06
Copyright © 2006, American Society for Microbiology. All Rights Reserved.



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