Purification and partial characterization of equine infectious anemia virus reverse transcriptase.

Abstract:

Previously we raised a rabbit monospecific antibody (C2003) against a synthetic peptide derived from a sequence within the C-terminal portion of the reverse transcriptase (RT) of the human immunodeficiency virus type 1 (HIV-1). This sequence is found to be conserved in the predicted amino acid sequence of a related lentivirus, the equine infectious anemia virus (EIAV). It was previously determined that the C2003 antibody could cross-react with native EIAV RT and directly inhibit the DNA polymerase activity of the enzyme. We have now fractionated EIAV RT by immunoaffinity chromatography with immobilized C2003 antibody. The procedure yielded an equimolar mixture of two proteins of 66 and 51 kDa associated with both DNA polymerase and RNase H activities. When the EIAV RT proteins were examined by in situ activity gel assays, polymerase activity was found to be principally associated with the 66-kDa component. The fidelity of DNA synthesis by EIAV RT was found to be equivalent to that of HIV-1 RT and lower than that of AMV RT. These observations indicate that the RTs of EIAV and HIV-1 share similar structural and functional properties.

Polymerases:

Topics:

Fidelity, RNase H Activity, Exonuclease Activity, Reverse Transcriptase, Source / Purification

Status:

new topics/pols set partial results complete validated

Results:

Polymerase Reference Property Result Context
EIAV RT DeVico A1991 Reverse Transcriptase Activity Yes
EIAV RT DeVico A1991 3-5' Exonuclease (proofreading) No
EIAV RT DeVico A1991 Nucleotide Substitution Rate 0.003115 errors/bp
EIAV RT DeVico A1991 Nucleotide Substitution Rate 0.001111 errors/bp
EIAV RT DeVico A1991 Full length or truncated Full length
EIAV RT DeVico A1991 RNase H Yes
AMV DeVico A1991 Reverse Transcriptase Activity Yes
AMV DeVico A1991 3-5' Exonuclease (proofreading) No
AMV DeVico A1991 Nucleotide Substitution Rate 0.002193 errors/bp
AMV DeVico A1991 Nucleotide Substitution Rate 0.000349 errors/bp
AMV DeVico A1991 Full length or truncated Full length
AMV DeVico A1991 RNase H Yes
HIV RT DeVico A1991 Reverse Transcriptase Activity Yes
HIV RT DeVico A1991 3-5' Exonuclease (proofreading) No
HIV RT DeVico A1991 Nucleotide Substitution Rate 0.003155 errors/bp
HIV RT DeVico A1991 Nucleotide Substitution Rate 0.001477 errors/bp
HIV RT DeVico A1991 Full length or truncated Full length
HIV RT DeVico A1991 RNase H Yes

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