Cellular Internalization Mechanism and Intracellular Trafficking ofFilamentous M13 Phages Displaying a Cell-Penetrating Transbody and TATPeptide

Collection with item attached
2012
Item details URL
http://open-repository.kisti.re.kr/cube/handle/open_repository/475788.do
DOI
10.1371/journal.pone.0051813
Title
Cellular Internalization Mechanism and Intracellular Trafficking ofFilamentous M13 Phages Displaying a Cell-Penetrating Transbody and TATPeptide
Description
The authors would like to thank Dr. Sang Jick Kim (KRIBB, Korea) forproviding the phagemid pDR-D1 vector and bacterial strain ER2738. Thiswork was supported by Mid-career Researcher Program (2010-0009760), thePriority Research Center Program (2012-0006687), and the Global FrontierProject grant (2012M3A6A4054252) grants from the National ResearchFoundation, funded by the Korea government (MEST). The funders had norole in study design, data collection and analysis, decision to publish,or preparation of the manuscript.
abstract
Cellular internalization of bacteriophage by surface-displayed cell penetrating peptides has been reported, though the underlying mechanism remains elusive. Here we describe in detail the internalization mechanism and intracellular trafficking and stability of filamentous M13 phages, the cellular entry of which is mediated by surface-displayed cell-penetrating light chain variable domain 3D8 VL transbody (3D8 VL-M13) or TAT peptide (TAT-M13). Recombinant 3D8 VL-M13 and TAT-M13 phages were efficiently internalized into living mammalian cells via physiologically relevant, energy-dependent endocytosis and were recovered from the cells in their infective form with the yield of 3D8 VL-M13 being higher (0.005 similar to 0.01%) than that of TAT-M13 (0.001 similar to 0.005%). Biochemical and genetic studies revealed that 3D8 VL-M13 was internalized principally by caveolae-mediated endocytosis via interaction with heparan sulfate proteoglycans as cell surface receptors, whereas TAT-M13 was internalized by clathrin- and caveolae-mediated endocytosis utilizing chondroitin sulfate proteoglycans as cell surface receptors, suggesting that phage internalization occurs by physiological endocytotic mechanism through specific cell surface receptors rather than non-specific transcytotic pathways. Internalized 3D8 VL-M13 phages routed to the cytosol and remained stable for more than 18 h without further trafficking to other subcellular compartments, whereas TAT-M13 phages routed to several subcellular compartments before being degraded in lysosomes even after 2 h of internalization. Our results suggest that the internalizing mechanism and intracellular trafficking of filamentous M13 bacteriophages largely follow the attributes of the displayed cell-penetrating moiety. Efficient internalization and cytosolic localization of 3D8 VL transbody-displayed phages will provide a useful tool for intracellular delivery of polar macromolecules such as proteins, peptides, and siRNAs. Citation: Kim
provenance
Made available in Cube on 2018-09-28T11:22:13Z (GMT). No. of bitstreams: 0
language
English
author
Kim, Aeyung
Shin, Tae-Hwan
Shin, Seung-Min
Pham, Chuong D.
Choi, Dong-Ki
Kwon, Myung-Hee
Kim, Yong-Sung
orcid
Pham, Dinh-Chuong/0000-0002-3074-6941; Kim,
Yong-Sung/0000-0003-2673-1509
accessioned
2018-09-28T11:22:13Z
available
2018-09-28T11:22:13Z
issued
2012
citation
PLOS ONE(7): 12
issn
1932-6203
uri
http://open-repository.kisti.re.kr/cube/handle/open_repository/475788.do
Funder
교육과학기술부
Funding Program
2단계연구중심대학육성(0.5)
Project ID
1345196741
Jurisdiction
Rep.of Korea
Project Name
Division of Cell Transformation and Restoration
rights
openAccess
type
article


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