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PMID: 10823846 Published · ppublish English Journal Article Research Support, Non-U.S. Gov't

Efficient DNA transfection mediated by the C-terminal domain of human immunodeficiency virus type 1 viral protein R.

Journal of virology ·Vol. 74 ·No. 12 ·2000-06-00 ·Pages 5424-31

Kichler A, Pages JC, Leborgne C, Druillennec S, Lenoir C, Coulaud D, Delain E, Le Cam E, Roques BP, Danos O

Abstract

Viral protein R (Vpr) of human immunodeficiency virus type 1 is produced late in the virus life cycle and is assembled into the virion through binding to the Gag protein. It is known to play a significant role early in the viral life cycle by facilitating the nuclear import of the preintegration complex in nondividing cells. Vpr is also able to interact with nucleic acids, and we show here that it induces condensation of plasmid DNA. We have explored the possibility of using these properties in DNA transfection experiments. We report that the C-terminal half of the protein (Vpr(52-96)) mediates DNA transfection in a variety of human and nonhuman cell lines with efficiencies comparable to those of the best-known transfection agents. Compared with polylysine, a standard polycationic transfection reagent, Vpr(52-96) was 10- to 1,000-fold more active. Vpr(52-96)-DNA complexes were able to reach the cell nucleus through a pH-independent mechanism. These observations possibly identify an alternate pathway for DNA transfection.

MeSH Terms
Amino Acid Sequence Animals Anti-Bacterial Agents/pharmacology Cell Cycle Cell Line Cell Nucleus/genetics,metabolism Chemical Precipitation Chloroquine/pharmacology DNA-Binding Proteins/chemistry,genetics,metabolism,ultrastructure Gene Products, vpr/chemistry,genetics,metabolism Genes, Reporter/genetics HIV-1 Humans Hydrogen-Ion Concentration Macrolides Mice Microscopy, Electron Molecular Sequence Data Peptide Fragments/chemistry,genetics,metabolism Plasmids/genetics,metabolism Polylysine/metabolism Promoter Regions, Genetic/genetics Protein Binding Transcriptional Activation/drug effects,genetics Transfection/methods vpr Gene Products, Human Immunodeficiency Virus
Chemicals
Anti-Bacterial Agents DNA-Binding Proteins Gene Products, vpr Macrolides Peptide Fragments vpr Gene Products, Human Immunodeficiency Virus Polylysine Chloroquine bafilomycin A1
Authors & Affiliations
10 authors, click to expand affiliations / ORCID
Kichler A
Généthon III, CNRS URA 1923, Evry, France. akich@genethon.fr
Pages J C
Leborgne C
Druillennec S
Lenoir C
Coulaud D
Delain E
Le Cam E
Roques B P
Danos O
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52 references, click to expand
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Article Info
Journal
Journal of virology
Abbr.
J Virol
ISSN
0022-538X
DOI
10.1128/jvi.74.12.5424-5431.2000
Email
akich@genethon.fr
Published
2000-06-00
Pages
5424-31
Language
English
Region
United States
NLM ID
0113724
PMCID
PMC112026
Subset
IM
Corrections
ErratumIn
-
External Links
PubMed source DOI: 10.1128/jvi.74.12.5424-5431.2000 PMC Full Text View journal details
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