Nucleic Acids Research, 1994, Vol. 22, No. 20 4125-4131
© 1994
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Characterization of the minimal DNA-binding domain of the HIV integrase protein
Division of Molecular Biology, The Netherlands Cancer Institute Plesmanlaan 121, 1066 CX Amsterdam, The Netherlands
*To whom correspondence should be addressed
Received July 6, 1994. Revised September 2, 1994. Accepted September 2, 1994.
The human immunodeficiency virus (HIV) integrase (IN) protein mediates an essential step in the retroviral lifecycle, the integration of viral DNA into human DNA. A DNA-binding domain of HIV IN has previously been identified in the C-terminal part of the protein. We tested truncated proteins of the C-terminal region of HIV-1 IN for DNA binding activity in two different assays: UV-crosslinking and southwestern blot analysis. We found that a polypeptide fragment of 50 amino acids (IN220–270) is sufficient for DNA binding. In contrast to full-length IN protein, this domain is soluble under low salt conditions. DNA binding of IN220–270 to both viral DNA and non-specific DNA occurs in an ion-independent fashion. Point mutations were introduced in 10 different amino acid residues of the DNA-binding domain of HIV-2 IN. Mutation of basic amino acid K264 results in strong reduction of DNA binding and of integrase activity.
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