Transient cell proliferation with polyethylenimine-cationized N-terminal domain of simian virus 40 large T-antigen.
Polyethylenimine (PEI) cationization is a powerful strategy for protein transduction into cells. In this study, we attempted the artificial regulation of cell proliferation by protein transduction of the N-terminal domain (1-132 amino acids) of the simian virus 40 large T-antigen (SVLT-N), which inactivates retinoblastoma family proteins but not p53. To deliver SVLT-N into cells, we employed an indirect cationization method by forming a complex of biotynylated SVLT-N through disulfide bonds (biotin-SS-SVLT-N) and PEI-cationized avidin (PEI600-avidin). Using this complex, SVLT-N was transduced into the nucleus of confluent and quiescent Balb/c 3T3 cells and was found to be complexed with a cellular target protein, pRb. Furthermore, SVLT-N transduction induced cell proliferation in spite of confluent conditions. Because SVLT-N thus transduced into cells gradually degraded and was not detectable after a 4-d incubation, transiently transformed cells were obtained by this method. These results suggest that oncogene protein transduction technology has great potential for in vitro regulation of cell proliferation.
Murata
Hitoshi
H
Department of Bioscience and Biotechnology, Faculty of Engineering, Graduate School of Natural Science and Technology, Okayama University, Okayama, Japan.
Futami
Junichiro
J
Kitazoe
Midori
M
Kosaka
Megumi
M
Tada
Hiroko
H
Seno
Masaharu
M
Yamada
Hidenori
H
D000818:Animals; D000952:Antigens, Polyomavirus Transforming / Q000737:chemistry* / Q000378:metabolism / Q000494:pharmacology*; D001360:Avidin / Q000737:chemistry*; D041702:BALB 3T3 Cells; D019921:Biotinylation; D049109:Cell Proliferation / Q000187:drug effects*; D008722:Methods; D051379:Mice; D011094:Polyethyleneimine / Q000737:chemistry*; D017434:Protein Structure, Tertiary; D021381:Protein Transport; D016160:Retinoblastoma Protein / Q000378:metabolism; D013539:Simian virus 40
D000952:Antigens, Polyomavirus Transforming; C528510:PEI600-avidin; D016160:Retinoblastoma Protein; D001360:Avidin; D011094:Polyethyleneimine