Parameters that influence processive synthesis and site-specific termination by human immunodeficiency virus reverse transcriptase on RNA and DNA templates.

Abstract:

We have examined the parameters that determine the length and distribution of products synthesized processively by the human immunodeficiency virus reverse transcriptase (HIV-RT). On native or homopolymer templates, the overall length distribution of processively synthesized products is increased by increased temperature or deoxynucleoside triphosphate concentration, or decreased ionic strength. Specific terminations of processive synthesis on either native DNA or RNA templates occur most frequently at positions where the reverse transcriptase (RT) pauses during synthesis. These sites correlate with the template sequence 3'-(A/U)(A/U)(G/C)-5', particularly when this sequence is predicted to be base paired with another region of the template in a secondary structure. Many positions of termination are in similar positions on DNA or RNA templates. Notable exceptions are runs of A residues, which promote termination on DNA but not RNA templates. Termination intensities vary when different RTs are used demonstrating an influence of RT structure.

Polymerases:

Topics:

Historical Protein Properties (MW, pI, ...), Source / Purification, RNase H Activity, Nucleotide Incorporation, Reverse Transcriptase

Status:

new topics/pols set partial results complete validated

Results:

Polymerase Reference Property Result Context
MMuLV RT DeStefano JJ1992 Reverse Transcriptase Activity Yes
MMuLV RT DeStefano JJ1992 Cloned or native Native organism
MMuLV RT DeStefano JJ1992 Full length or truncated Full length
MMuLV RT DeStefano JJ1992 RNase H Yes
MMuLV RT DeStefano JJ1992 Specific Activity 1E+05 units/mg
HIV RT DeStefano JJ1992 Reverse Transcriptase Activity Yes
HIV RT DeStefano JJ1992 Molecular Weight 1.17E+05 Dalton
HIV RT DeStefano JJ1992 Full length or truncated Full length
HIV RT DeStefano JJ1992 RNase H Yes
HIV RT DeStefano JJ1992 Processivity 75bp
HIV RT DeStefano JJ1992 Specific Activity 2.1E+04 units/mg
AMV DeStefano JJ1992 Reverse Transcriptase Activity Yes
AMV DeStefano JJ1992 Cloned or native Native organism
AMV DeStefano JJ1992 Tagged No
AMV DeStefano JJ1992 Full length or truncated Full length
AMV DeStefano JJ1992 RNase H Yes
AMV DeStefano JJ1992 Specific Activity 6.2E+04 units/mg

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