A long synthetic peptide containing a nuclear localization signal and its flanking sequences of SV40 T-antigen directs the transport of IgM into the nucleus efficiently

Yoneda, Yoshihiro; Semba, Toshihiko; Kaneda, Y.; Noble, Richard L.; Matsuoka, Yosuke; Kurihara, Takushi; Okada, Yoshio; Imamoto, Naoko

doi:10.1016/0014-4827(92)90279-h

Cited by 58 publications

(26 citation statements)

References 33 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…NLS-phage displays a 32-mer peptide derived from the SV40 T-antigen (Fig. 1A), which functions as a strong nuclear localization signal (20). In a separate study, we found that NLS-phage can actively penetrate into the nucleus when injected directly into the cytoplasm.…”

Section: Resultsmentioning

confidence: 79%

“…Preparation of Recombinant Phage Particles-The protein transduction domain of Tat protein (Tat peptide) (11), the integrin-binding peptide (RGD peptide) (18), the heparin-binding domain of vitronectin (VN peptide) (19), and the nuclear localization signal (NLS) of simian virus 40 (SV40) T antigen (20) were displayed on phage heads, as described previously (21), except that we used our original lysogenic bacterial strains. D1180 (Dam15 del EcoRI-SacI cIts857 nin5 Sam100) was generated by crossing Dam15 (22) and gt11 (23), followed by deletion between unique EcoRI and SacI sites by linker oligonucleotides.…”

Section: Methodsmentioning

confidence: 99%

See 1 more Smart Citation

Protein Transduction Domain of HIV-1 Tat Protein Promotes Efficient Delivery of DNA into Mammalian Cells

Eguchi¹,

Akuta²,

Okuyama³

et al. 2001

Journal of Biological Chemistry

280

296

View full text Add to dashboard Cite

The plasma membrane of mammalian cells is one of the tight barriers against gene transfer by synthetic delivery systems. Various agents have been used to facilitate gene transfer by destabilizing the endosomal membrane under acidic conditions, but their utility is limited, especially for gene transfer in vivo. In this article, we report that the protein transduction domain of human immunodeficiency virus type 1 Tat protein (Tat peptide) greatly facilitates gene transfer via membrane destabilization. We constructed recombinant phage particles displaying Tat peptide on their surfaces and carrying mammalian marker genes as part of their genomes (Tat-phage). We demonstrate that, when animal cells are briefly exposed to Tat-phage, significant expression of phage marker genes is induced with no harmful effects to the cells. In contrast, recombinant phage displaying other functional peptides, such as the integrin-binding domain or a nuclear localization signal, could not induce detectable marker gene expression. The expression of marker genes induced by Tatphage is not affected by endosomotropic agents but is partially impaired by inhibitors of caveolae formation. These data suggest that Tat peptide will become a useful component of synthetic delivery vehicles that promote gene transfer independently of the classical endocytic pathway.

show abstract

Section: Resultsmentioning

confidence: 79%

Section: Methodsmentioning

confidence: 99%

Protein Transduction Domain of HIV-1 Tat Protein Promotes Efficient Delivery of DNA into Mammalian Cells

Eguchi¹,

Akuta²,

Okuyama³

et al. 2001

Journal of Biological Chemistry

280

296

View full text Add to dashboard Cite

show abstract

“…The NLS peptides coupled by way of covalent linkers have already been demonstrated to confer nuclear localization on a variety of proteins, e.g., BSA and ferritin (Lanford et al, 1986), and even molecules as large as IgM (Yoneda et al, 1992). It is therefore conceivable that NLSs could be harnessed to improve the delivery of DNA, but it is not yet clear how best to incorporate the NLS peptides into gene delivery systems.…”

Section: Use Of Nls Pathways For Nuclear Import Of Gene Delivery Vectorsmentioning

confidence: 99%

Improvement of exogenous DNA nuclear importation by nuclear localization signal‐bearing vectors: a promising way for non‐viral gene therapy?

Hebert

2003

Biology of the Cell

View full text Add to dashboard Cite

Several vectors have been developed in order to target genes to specific cells. Virus-based vectors lead to a high transfection efficiency in vitro, but display important disadvantages such as pathological risks, which they expose to patients. Plasmid-associated chemical vectors lack these disadvantages, but allow only a very low efficiency of transgene expression. Most of the non-viral-based gene transfer techniques developed until now mainly focused their efforts to overcome the problem of DNA entry into the cell. Some recent works, however, have begun to investigate the nucleus entry problem and suggest that the trafficking of DNA from cytosol to the nucleus may be improved by using the nuclear localization signal (NLS) found in some nuclear proteins. If the vector contains one or several NLS, either as covalently or non-covalently DNA-linked peptides, a competition may take place between the rate of dissociation of the DNA-vector complexes and the rate of loading of the complexes to the NLS-mediated nucleus importation machinery. This equilibrium may be displaced towards the importation pathway by the use of NLS-bearing proteins instead of peptides. The possibility of recruiting normal endogenous cellular pathways of nuclear uptake to promote entry of exogenously applied DNA through the nuclear pore complex would, thus, seem promising. Nevertheless, attempts to improve the transport of DNA to the nucleus through the use of NLSs have achieved limited success. Although these systems show improved transgene expression, little is known about how they function in transfected cells, and the optimal formulation for gene expression is yet to be determined.

show abstract

“…This construct was called scFvCk and expressed in E. coli to check its ability to bind to human p53 (data not shown). For nuclear targeting, the scFv and scFvCk were additionally modi®ed to contain the C-terminal SV40 nuclear localization signal TPPKKKRKV which has been shown to direct heterologous proteins into the nucleus (Yoneda et al, 1992). The two nucleus-targeted scFvs were called respectively scFv-SV40 (Figure 1b) and scFvCk-SV40 (Figure 1d).…”

Section: Do-1 Scfv Is a Powerful Protecting Agent Of Dna Binding Actimentioning

confidence: 99%

Characterization of a new intrabody directed against the N-terminal region of human p53

1998

View full text Add to dashboard Cite

Genes encoding the rearranged immunoglobulin heavy and light chain variable regions of DO-1, a monoclonal antibody directed against human p53, have been used to construct a single-chain antibody. DO-1 recognizes an N-terminal epitope in the region involved in the transactivation function of p53 and the binding of Mdm2. The DO-1 single chain scFv expressed in the periplasm of E. coli or at the surface of the ®lamentous phage M13 retained the immunological speci®city and a nity of the full length antibody. Furthermore, the DO-1 recombinant antibody was able to inhibit the in vitro binding of Hdm2, and was shown to be a powerful protecting agent of p53's DNA binding activity at 378C. The DO-1 single-chain antibody has been used to construct single-chain intracellular antibodies (intrabodies) for expression in the cytoplasm and the nucleus of mammalian cells. These anti-p53 intrabodies were additionally modi®ed by addition of a Ck domain to increase cytoplasmic and nuclear stability. Here we show that expression of the DO-1 single-chain antibody in the H1299 cell line results in an inhibition of p53's transactivation function. The DO-1 intrabody is a useful tool to study those functions of p53 driven by the Nterminal region of the protein.

show abstract

A long synthetic peptide containing a nuclear localization signal and its flanking sequences of SV40 T-antigen directs the transport of IgM into the nucleus efficiently

Cited by 58 publications

References 33 publications

Protein Transduction Domain of HIV-1 Tat Protein Promotes Efficient Delivery of DNA into Mammalian Cells

Protein Transduction Domain of HIV-1 Tat Protein Promotes Efficient Delivery of DNA into Mammalian Cells

Improvement of exogenous DNA nuclear importation by nuclear localization signal‐bearing vectors: a promising way for non‐viral gene therapy?

Characterization of a new intrabody directed against the N-terminal region of human p53

Contact Info

Product

Resources

About