Fusion protein comprising tat-derived transport moiety

Details Fusion protein comprising tat-derived transport moiety Fusion protein comprising tat-derived transport moiety

1995-05-25

1997-10-07

Elliott, George C.

Chemistry: natural resins or derivatives; peptides or proteins;

Proteins, i.e., more than 100 amino acid residues

C07K 1900, C07K 1400, C07K 14005

Patent

active

056749805

ABSTRACT:
This invention relates to delivery of biologically active cargo molecules, such as polypeptides and nucleic acids, into the cytoplasm and nuclei of cells in vitro and in vivo. Intracellular delivery of cargo molecules according to this invention is accomplished by the use of novel transport polypeptides which comprise HIV tat protein or one or more portions thereof, and which are covalently attached to cargo molecules. The transport polypeptides in preferred embodiments of this invention are characterized by the presence of the tat basic region (amino acids 49-57), the absence of the tat cysteine-rich region (amino acids 22-36) and the absence of the tat exon 2-encoded carboxy-terminal domain (amino acids 73-86) of the naturally-occurring tat protein. By virtue of the absence of the cysteine-rich region, the preferred transport polypeptides of this invention solve the potential problems of spurious trans-activation and disulfide aggregation. The reduced size of the preferred transport polypeptides of this invention also minimizes interference with the biological activity of the cargo molecule.

REFERENCES:
patent: 4861588 (1989-08-01), Neurath et al.
patent: 4918166 (1990-04-01), Kingsman et al.
patent: 5132213 (1992-07-01), Bachmair et al.
patent: 5219990 (1993-06-01), Androphy et al.
E.J. Androphy et al., "Bovine Papillomavirus E2-Trans-Activating Gene Product Binds to Specific Sites in Papillomavirus DNA", Nature, 325, pp. 70-73 (1987).
S.K. Arya et al., "Trans-Activator Gene of Human T-Lymphotropic Virus Type III (HTLV-III)", Science, 229:69-73 (1985).
S.K. Arya and R.C. Gallo, "Three Novel Genes of Human T-Lymphotropic Virus Type III: Immune Reactivity of their Products with Sera from Acquired Immune Deficiency Syndrome Patients", Proc. Natl. Acad. Sci. USA, 83:2209-13 (1986).
S.K. Arya et al., "New Human and Simian HIV-Related Retroviruses Possess Functional Transactivator (tat) Gene", Nature, vol. 328, pp. 548-550 (1987).
J. Barsoum, "Introduction of Stable High-Copy-Number DNA into Chinese Hamster Ovary Cells by Electroporation", DNA Cell Biol., 9, pp. 293-300 (1990).
J. Barsoum et al., "Mechanism of Action of the Papillomavirus E2 Repressor: Repression in the Absence of DNA Binding", J. Virol., vol. 66, pp. 3941-3945 (1992).
R. Carroll et al., "Identification of Lentivirus Tat Functional Domains through Generation of Equine Infectious Anemia Virus/Human Immunodeficiency Virus Type 1 tat Gene Chimeras", J. Virol., vol., 65, pp. 3460-3467 (1991).
R. Carroll et al., "Inhibition of Human Immunedeficiency Virus Type 1 Tat Activity by Coexpreesion of Heterologous trans Activators", J. Virol., vol., 66, pp. 2000-2007 (1992).
R.L. Cate et al., "Isolation of the Bovine and Human Genes for Mullerian Inhibiting Substance and Expression of the Human Gene in Animal Cells", Cell, vol. 45, pp. 685-698 (1986).
L. Chakrabarti et al., "Sequence of Simian Immunodeficiency Virus from Macaque and Its Relationship to other Human and Simian Retroviruses", Nature, vol. 328, pp. 543-547 (1987).
J. Choe et al., "Bovine Papillomavirus Type 1 Encodes Two Forms of a Transcriptional Repressor: Structural and Functional Analysis of New Viral cDNAs", J. Virol., 63, pp. 1743-1755 (1989).
G. Chu et al., "Electroporation for the Efficient Transfection of Mammalian Cells with DNA", Nuc. Acids Res., 15, pp. 1311-1326 (1987).
B.R. Cullen, "Trans-Activation of Human Immunedeficiency Virus Occurs Via A Bimodal Mechanism", Cell, 46:973-982 (1986).
C.V. Dang and W.M.F. Lee, "Nuclear and Nucleolar Targeting Sequences of c-erb-A, c-myb, N-myc, p53, HSP70, and HIV tat Proteins", J.Biol.Chem., 264:18019-23 (1989).
N. Dostatni et al., "A Dimer of BPV-1 E2 Containing a Protease Resistant Core Interacts with its DNA Target", EMBO J., 7, pp. 3807-3816 (1988).
H. Durst et al., "A Papillomavirus DNA from a Cervical Carcinoma and Its prevalence in Cancer Biopsy Samples from Different Geographic Regions", Proc. Natl. Acad. Sci. USA, vol. 80, pp. 3812-3815 (1983).
B. Elangovan et al., "Functional Comparison of the Basic Domains of the Tat Proteins of Human Immunodeficiency Virus Types 1 and 2 in trans Activation", J. Virol., vol., 66, pp. 2031-2036 (1992).
M. Emerman et al., "The Specificity of the Human Immunodeficiency Virus Type 2 Transactivator Is Different From That of Human Immunodeficiency Virus Type 1", EMBO J., vol. 6, pp. 3755-3760 (1987).
H. Farhood et al., "Regulated Gene Transfer By Co-Delivery Of A Cis-Acting DNA Element And A Trans-Acting Protein Factor To Mammalian Cells With Cationic Liposomes", J. Cell. Biochem., Supp 17E:242 (1993).
R. Fenrick et al., "Functional Analysis of the Tat trans Activator of Human Immunodeficiency Virus Type 2", J. Virol., vol., 63, pp. 5006-5012 (1989).
A.D. Frankel et al., "Activity of Synthetic Peptides from the Tat Protein of Human Immunodeficiency Virus Type 1," Proc. Natl. Acad. Sci. USA, 86:7397-401 (1989).
A.D. Frankel and C.O. Pabo, "Cellular Uptake of the Tat Protein from Human Immunodeficiency Virus", Cell, 55:1189-93 (1988).
A.D. Frankel et al., "Tat Protein from Human Immunodeficiency Virus Forms a Metal-Linked Dimer," Science, 240:70-73 (1988).
A.D. Frankel et al., "Dimerization of the Tat Protein from Human Immunodeficiency Virus: A Cysteine-Rich Peptide Mimics the Normal Metal-Linked Dimer Interface", Proc. Natl. Acad. Sci. USA, 85:6297-300 (1988).
J.A. Garcia et al., "Functional Domains Required for Tat-induced Transcriptional Activation of the HIV-1 Long Terminal Repeat", EMBO J., 7:3143-47 (1988).
I. Giri et al., "Study of the E2 Gene Product of the Cottontail Rabbit Papillomavirus Reveals a Common Mechanism of Transactivation Among Papillomaviruses", J. Virol., 62, pp. 1573-1581 (1988b).
I. Giri et al., "Structural and Mutational Analysis of E2 Trans-Activating Proteins of Papillomaviruses Reveals Three Distinct Functional Domains", EMBO J., 7, pp. 2823-2829 (1988a).
M. Green and P.M. Loewenstein, "Autonomous Functional Domains of Chemically Synthesized Human Immunodeficiency Virus Tat Trans-Activator Protein", Cell, 55:1179-88 (1988).
M. Green et al., "Mutationa1 Analysis of HIV-1 Tat Minimal Domain Peptides: Identification of Trans-Dominant Mutants That Suppress HIV-LTR-Driven Gene Expression", Cell, 58:215-23 (1989).
M. Guyader et al., "Genome Organization and Transactivation of the Human Immunodeficiency Virus Type 2", Nature, vol. 326, pp. 662-669 (1987).
G.S. Harrison et al., "Toward HIV-Regulated Expression of A Diptheria Toxin A Gene In Transfected Cells", J. Cell. Biochem., Supp. 13B:302 (1989).
J. Hauber et al,, "Mutationa1 Analysis of the Conserved Basic Domain of Human Immunodeficiency Virus tat Protein," J. Virol., 63:1181-87 (1989).
T.H. Haugen et al. "Trans-Activation of an Upstream Early Gene Promoter of Bovine Papillomavirus-1 by a Product of the Vital E2 Gene", EMBO J., 6, pp. 145-152 (1987).
T.H. Haugen et al., "Sequence-Specific and General Transcriptional Activation by the Bovine Papillomavirus-1 E2 trans-Activator Require an N-Terminal Amphipathic Helix-Containing E2 Domain", EMBO J., 7. pp. 4245-4253 (1988).
P. Hawley-Nelson et al., "The Specific DNA Recognition Sequence of the Bovine Papillomavirus E2 Protein is an E2-Dependent Enhancer", EMBO J., 7, pp. 525-531 (1988).
H. Hirochika et al., "Enhancers and trans-Acting E2 Transcriptional Factors of Papillomaviruses", J. Virol., 61, pp. 2599-2606 (1987).
H. Hirochika et al., "Functional Mapping of the Human Papillomavirus Type 11 Transcriptional Enhancer and its Interaction with the trans-acting E2 Proteins", Genes Dev., 2, 54-67 (1988).
V. Hirsch et al., "The Genome Organization of STLV-3 is Similar to That of the AIDS Virus Except for a Truncated Transmembrane Protein", Cell, vol. 49, pp. 307-319 (1987).
P. Hoffmann et al., "Stimulation of Human and Murine Adherent Cells by Bacterial LIpoprotein and Synthetic Llpopeptide Analoguss", Immunobiol., vol. 177, pp. 158-70 (1988).
D. Kalderon et al., "A Short Amino Acid Sequence Able to Specify Nuclear Location," Cell, 39:499-509 (1984).
M. Kuppuswamy et al., "Multiple Functional Domains of Tat, the Trans-activator of HIV-1, Defined by Muta

Affiliated with

Also associated with

Rating

Fusion protein comprising tat-derived transport moiety does not yet have a rating. At this time, there are no reviews or comments for this patent.

If you have personal experience with Fusion protein comprising tat-derived transport moiety, we encourage you to share that experience with our LandOfFree.com community. Your opinion is very important and Fusion protein comprising tat-derived transport moiety will most certainly appreciate the feedback.

Rate now

Comments { 0 }

Profile ID: LFUS-PAI-O-2358268