The K65R mutation confers increased DNA polymerase processivity to HIV-1 reverse transcriptase.

Abstract:

The K65R mutation in HIV-1 reverse transcriptase (RT) is associated ...
The K65R mutation in HIV-1 reverse transcriptase (RT) is associated with viral cross-resistance to 2',3'-dideoxyinosine, 2',3'-dideoxycytidine, and 2',3'-dideoxy-3'-thiacytidine. We have found that in vitro DNA synthesis by K65R RT is significantly more processive than that of wild type (wt) RT. Depending on the template/primer (T/P) used, the total incorporation of nucleotides under single processive cycle conditions was 20-50% higher with K65R RT than with wt RT. With heteropolymeric T/P, the total incorporation of dNMP by K65R and wt RT was similar under continuous DNA synthesis reaction conditions. However, under single processive cycle conditions, the rate of full-length polymerization product synthesis by K65R RT was about 2-fold higher than that by wt RT. We also found a decreased rate of T/P dissociation during K65R RT DNA synthesis, which is consistent with the increased processivity of the enzyme. We postulate that the increased processivity of the K65R RT may be a compensatory response to the decreased affinity of this mutant for certain dNTP substrates, allowing normal viral replication kinetics.

Polymerases:

Topics:

Other Enzymatic Activities, Reverse Transcriptase, Source / Purification

Status:

new topics/pols set partial results complete validated

Results:

Polymerase Reference Property Result Context
HIV RT The K65R mutation confers increased DNA polymerase processivity to HIV-1 reverse transcriptase. Reverse Transcriptase Activity Yes
HIV RT The K65R mutation confers increased DNA polymerase processivity to HIV-1 reverse transcriptase. Cloned or native Cloned in E. coli
HIV RT The K65R mutation confers increased DNA polymerase processivity to HIV-1 reverse transcriptase. Full length or truncated Full length
HIV RT The K65R mutation confers increased DNA polymerase processivity to HIV-1 reverse transcriptase. Extension from RNA primer Yes
HIV RT K65R The K65R mutation confers increased DNA polymerase processivity to HIV-1 reverse transcriptase. Reverse Transcriptase Activity Yes
HIV RT K65R The K65R mutation confers increased DNA polymerase processivity to HIV-1 reverse transcriptase. Cloned or native Cloned in E. coli
HIV RT K65R The K65R mutation confers increased DNA polymerase processivity to HIV-1 reverse transcriptase. Full length or truncated Full length
HIV RT K65R The K65R mutation confers increased DNA polymerase processivity to HIV-1 reverse transcriptase. Extension from RNA primer Yes

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