Polynucleotides encoding a human TRIM-Cyp fusion polypeptide, compositions thereof, and methods of using same
A nucleic acid is provided which encodes a human TRIM-cyclophilin A fusion sequence encoding a human TRIM-CypA fusion protein which is active as an anti-viral agent, and in particular, as an anti-HIV-1 agent. Also provided is a nucleic acid encoding a polypeptide having both TRIM activity and cyclophilin activity. Also provided is an isolated polynucleotide encoding a human TRIM-CypA fusion protein, or variants thereof retaining the TRIM and cyclophilin A activities. Also provided are compositions thereof, antibodies that specifically bind thereto, and vectors and host cells comprising the nucleic acid or polypeptide. In addition, methods are provided for treating or preventing viral infection, or reducing viral load in a subject comprising administering the nucleic acid, polypeptide, vector, or composition to the subject in an amount effective to treat or prevent the viral infection. In some embodiments, the viral infection is HIV-I infection, hepatitis C infection, pox virus infection, vaccinia virus infection, or HTLV infection.
1. A nucleic acid which comprises a sequence selected from the group consisting of SEQ ID NO:5, SEQ ID NO:6, SEQ ID NO:7, a variant having at least 87% identity to SEQ ID NO:5, a variant having at least 94% identity to SEQ ID NO:6, and a variant having at least 89% identity to SEQ ID NO:7.
2. A nucleic acid which comprises a sequence complementary to the nucleic acid of claim 1 .
3. A nucleic acid encoding a polypeptide comprising SEQ ID NO:10, 11 or 12.
4. The nucleic acid of claim 1 , encoding a human TRIM5-cyclophilin A fusion protein which comprises a polypeptide linker sequence, wherein (i) TRIM5 is located upstream of the linker sequence, so that the N-terminal amino acid of the linker sequence is attached to the C-terminal amino acid of TRIM5, and (ii) cyclophilin A is located downstream of the linker sequence, so that the C-terminal amino acid residue of the linker sequence is attached to the N-terminal amino acid residue of cyclophilin A.
5. The nucleic acid of claim 4 , wherein the linker sequence comprises from about 10 to about 20 amino acids.
6. A polypeptide encoded by the nucleic acid of any of claim 1 , 2 , 3 , 4 , or 5 .
7. A nucleic acid vector, which comprises the nucleic acid of any one of claim 1 , 2 , 3 , 4 , or 5 .
8. A composition comprising the nucleic acid of claim 1 , 2 , 3 , 4 or 5 , and a carrier.
9. An isolated host cell comprising the nucleic acid of claim 1 , 2 , 3 , 4 , or 5 .
10. The nucleic acid of claim 1 , wherein the variant has at least 89%, 91%, 93%, 95%, or 99% identity to SEQ ID NO:5.
11. The nucleic acid of claim 1 , wherein the variant has at least 95%, 96%, 97%, 98%, or 99% identity to SEQ ID NO:6.
12. The nucleic acid of claim 1 , wherein the variant has at least 90%, 92%, 94%, 96%, or 98% identity to SEQ ID NO:7.