Use of the baculovirus system to assemble polyomavirus capsid-like particles with different polyomavirus structural proteins: analysis of the recombinant assembled capsid-like particles.

An, Ke; Gillock, Eric T.; Sweat, Jeffrey A.; Reeves, Wendy; Consigli, Richard A.

doi:10.1099/0022-1317-80-4-1009

Cited by 32 publications

(13 citation statements)

References 42 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…Py-VLPs expressed in insect cells frequently contain cellular DNA (1,20). Analysis of the partially purified Py-VLPs derived from VP1/wt, VP1/HI-FLAG, and VP1/EF-uPA(1-60)/ HI-FLAGϩVP1/HI-FLAG by agarose gel electrophoresis after DNase I treatment showed that they contained ϳ5 kb of FIG.…”

Section: Resultsmentioning

confidence: 99%

Formation of Polyomavirus-Like Particles with Different VP1 Molecules That Bind the Urokinase Plasminogen Activator Receptor

Shin

Folk

2003

J Virol

View full text Add to dashboard Cite

Icosahedral virus-like particles formed by the self-assembly of polyomavirus capsid proteins (Py-VLPs) can serve as useful nanostructures for delivering nucleic acids, proteins, and pharmaceuticals into animal cells and tissues. Four predominant surface-exposed loops in the VP1 structure offer potential sites to display sequences that might contribute new targeting specificities. Introduction into each of these loops of sequences derived from the amino-terminal fragment of urokinase plasminogen activator (uPA) or a related phage display peptide reduced the solubility of VP1 molecules when expressed in insect cells, and insertions into the EF loop reduced VP1 solubility least. Coexpression in insect cells of the uPA-VP1 molecules and VP1 containing a FLAG epitope in the HI loop permitted the formation of heterotypic Py-VLPs containing uPA-VP1 and FLAG-VP1. These heterotypic VLPs bound to uPAR on the surfaces of animal cells. Heterotypic Py-VLPs containing ligands for multiple cell surface receptors should be useful for targeting specific cells and tissues.The polyomavirus virion is composed of 72 pentameric subunits arranged in an icosahedral lattice with an approximate diameter of 50 nm (51). Polyomavirus virus-like particles (PyVLPs) of essentially the same dimensions and shape are formed by the self-assembly of the major capsid protein VP1 (55, 56). The structural framework for VP1 is an antiparallel ␤-sheet with jelly roll topology from which emanate four loops (58-60). The BC, DE, and HI loops closely interact and are located at the outward-facing end of the ␤-sheet; the EF loop, originating from the inward-facing end, is located on a side of the ␤-sheet (59, 60). A shallow groove formed between the BC1 and HI loops binds oligosaccharides terminating in ␣-2,3-linked sialic acid, expressed on many animal cells (8,18,58,60); the EF loop, together with the DE loop, contains motifs for integrin binding, which is important for cell susceptibility at a postattachment level (6). Among different strains of polyomavirus, the four VP1 surface-exposed loops exhibit sequence variability (3,17,36,39,53) and the HI loop has been used predominantly as a site for insertion so as to display on the surface of Py-VLPs the Escherichia coli dihydrofolate reductase (DHFR) (22), protein Z (21), a WW domain peptide (57), and a polyanionic adapter sequence (62).The urokinase plasminogen activator receptor (uPAR) is a glycosylphosphatidylinositol-linked protein expressed on the apical surfaces of endothelial cells (46) and certain epithelial cells (26) and leukocytes (48) to promote proteolysis (13), cell adhesion (7, 66), migration (5, 15), and chemotaxis (9,14,27,28,42,52). uPAR is expressed by many cancer cells, where it is correlated with metastasis and poor prognosis (10, 29-32, 43, 63), and is a potential target for drug and gene therapy. Also, uPAR is expressed on the apical surface of human airway epithelia and might be used to target delivery of DNAs to correct the genetic defect in individuals with cystic fibrosis (12). Se...

show abstract

Section: Resultsmentioning

confidence: 99%

Formation of Polyomavirus-Like Particles with Different VP1 Molecules That Bind the Urokinase Plasminogen Activator Receptor

Shin

Folk

2003

J Virol

View full text Add to dashboard Cite

show abstract

“…The combined abilities of the Py VP1 protein to self-assemble into virus-like particles (VLPs) and to nonspecifically interact with homologous or heterologous DNA has led to VP1 VLPs being proposed as potential vectors for gene therapy (1,35,51). VLPs efficiently transfer DNA and allow its expression in mammalian cells in vitro and in vivo with a wide tissue distribution (13,22,47).…”

Section: Discussionmentioning

confidence: 99%

α4β1 Integrin Acts as a Cell Receptor for Murine Polyomavirus at the Postattachment Level

Caruso

Belloni

Sthandier

et al. 2003

J Virol

View full text Add to dashboard Cite

The initial interaction of murine polyomavirus (Py) with host cells occurs through direct binding of the major capsid protein VP1 with cell membrane molecules containing terminal sialic acids; however, these Py receptor molecules have not yet been identified. Analysis of the capsid protein primary sequences of all murine strains revealed the presence of integrin ligand motifs in the DE and EF loops of VP1 (LDV and DLXXL, respectively) and at the N terminus of VP2 (DGE). We show that infectivity of the Py A2 strain in mouse Swiss 3T3 fibroblasts is significantly reduced only in the presence of natural integrin ligands carrying an LDV motif or antibodies directed against the ␣4 and ␤1 integrin subunits. Furthermore, we demonstrate that expression of the ␣4 subunit in the ␣4-deficient BALB/c 3T3 cells increases viral infectivity. Addition of ␣4 functionblocking antibodies, prior to or after virus adsorption, blocks this increased infectivity without affecting virus binding to cells. Taken together, these data indicate that expression of ␣4 integrin enhances permissivity to Py, probably by acting as one of the postattachment receptors.

show abstract

“…In support of this notion, pseudocapsids made from VP1 and VP2 are more effective in blocking interaction of virus with the cell, than those composed of VP1 alone. 39 Thus, a particular function may be missing from pseudocapsids, which, if replaced, could restore the efficiency of transfer to levels nearer that achieved with viruses.…”

Section: % Triton X 100 and Pseudocapsid Uptake Followed In Cells Bymentioning

confidence: 99%

Virus-like gene transfer into cells mediated by polyoma virus pseudocapsids

et al. 2000

View full text Add to dashboard Cite

Mouse polyoma virus-like particles (or pseudocapsids) are composed solely of recombinant viral coat protein. They can interact with DNA and transport it to cells, resulting in gene expression both in tissue culture and in mice. We demonstrate that DNA transfer in vitro depends on partial packaging of DNA within the virus-like capsid. Cell surface sialic acid residues and an intact microtubule network, required for viral infectivity, are also necessary for pseudocapsidmediated gene expression from heterologous DNA. Thus, gene delivery in this system requires pathways utilised by polyoma virions, rather than proceeding via the 'nonspecific' endosomal route typical of nonviral systems such as lipo-

show abstract

Use of the baculovirus system to assemble polyomavirus capsid-like particles with different polyomavirus structural proteins: analysis of the recombinant assembled capsid-like particles.

Cited by 32 publications

References 42 publications

Formation of Polyomavirus-Like Particles with Different VP1 Molecules That Bind the Urokinase Plasminogen Activator Receptor

Formation of Polyomavirus-Like Particles with Different VP1 Molecules That Bind the Urokinase Plasminogen Activator Receptor

α4β1 Integrin Acts as a Cell Receptor for Murine Polyomavirus at the Postattachment Level

Virus-like gene transfer into cells mediated by polyoma virus pseudocapsids

Contact Info

Product

Resources

About