Gene delivery of l-caldesmon protects cytoskeletal cell membrane integrity against adenovirus infection independently of myosin ATPase and actin assembly

Kari Haxhinasto; Anant Kamath; Ken Blackwell; James Bodmer; Jon Van Heukelom; Anthony English; Er-Wei Bai; Alan B Moy

doi:10.1152/ajpcell.00530.2003

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Gene delivery of l-caldesmon protects cytoskeletal cell membrane integrity against adenovirus infection independently of myosin ATPase and actin assembly

Journal article

Peer reviewed

Gene delivery of l-caldesmon protects cytoskeletal cell membrane integrity against adenovirus infection independently of myosin ATPase and actin assembly

Kari Haxhinasto, Anant Kamath, Ken Blackwell, James Bodmer, Jon Van Heukelom, Anthony English, Er-Wei Bai and Alan B Moy

American Journal of Physiology: Cell Physiology, Vol.287(4), pp.C1125-1138

10/2004

DOI: 10.1152/ajpcell.00530.2003

PMID: 15189814

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Abstract

The cytoskeleton is critical to the viral life cycle. Agents like cytochalasin inhibit viral infections but cannot be used for antiviral therapy because of their toxicity. We report the efficacy, safety, and mechanisms by which gene delivery of human wild-type low-molecular-weight caldesmon (l-CaD) protects cell membrane integrity from adenovirus infection in a DF-1 cell line, an immortalized avian fibroblast that is null for l-CaD. Transfection with an adenovirus (Ad)-controlled construct mediated a dose-dependent decline in transcellular resistance. In accordance with a computational model of cytoskeletal membrane properties, Ad disturbed cell-cell and cell-matrix adhesion and membrane capacitance. Transfection with the Ad-l-CaD construct attenuated adenovirus-mediated loss in transcellular resistance. Quantitation of vinculin-stained plaques revealed an increase in total focal contact mass in monolayers transfected with the Ad-l-CaD construct. Expression of l-CaD protected transcellular resistance through primary effects on membrane capacitance and independently of actin solubility and effects on pre-stress, as measured by the decline in isometric tension in response to cytochalasin D. Expression of l-CaD exhibited less Trypan blue cell toxicity than cytochalasin, and, unlike cytochalasin, it did not interfere with wound closure or adversely effect transcellular resistance. These findings demonstrate the gene delivery of wild-type human l-CaD as a potentially efficacious and safe agent that inhibits some of the cytopathic effects of adenovirus.

Calmodulin-Binding Proteins - genetics

Fibroblasts - virology

Adenoviridae Infections - pathology

Cell Line

Gene Transfer Techniques

Genetic Therapy

Adenoviridae Infections - therapy

Humans

Actins - metabolism

Calmodulin-Binding Proteins - metabolism

Adenoviridae - pathogenicity

Cell Membrane - physiology

Reverse Transcriptase Polymerase Chain Reaction

Animals

Transfection

Cell Adhesion - physiology

Cytoskeleton - virology

Models, Biological

Electric Capacitance

Cell Membrane - virology

Myosins - metabolism

Cytoskeleton - physiology

Fibroblasts - metabolism

Details

Title: Subtitle: Gene delivery of l-caldesmon protects cytoskeletal cell membrane integrity against adenovirus infection independently of myosin ATPase and actin assembly
Creators: Kari Haxhinasto - Department of Internal Medicine, C33 GH, University of Iowa College of Medicine, Iowa City, IA 52242, USA
Anant Kamath
Ken Blackwell
James Bodmer
Jon Van Heukelom
Anthony English
Er-Wei Bai
Alan B Moy
Resource Type: Journal article
Publication Details: American Journal of Physiology: Cell Physiology, Vol.287(4), pp.C1125-1138
DOI: 10.1152/ajpcell.00530.2003
PMID: 15189814
ISSN: 0363-6143
eISSN: 1522-1563
Grant note: GM-61732 / NIGMS NIH HHS
Language: English
Date published: 10/2004
Academic Unit: Roy J. Carver Department of Biomedical Engineering; Electrical and Computer Engineering; Emergency Medicine
Record Identifier: 9984083894202771

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