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Literature summary for 3.1.26.13 extracted from

  • Slack, R.L.; Spiriti, J.; Ahn, J.; Parniak, M.A.; Zuckerman, D.M.; Ishima, R.
    Structural integrity of the ribonuclease H domain in HIV-1 reverse transcriptase (2015), Proteins, 83, 1526-1538 .
    View publication on PubMedView publication on EuropePMC
Search for more literature for 3.1.26.13

Cloned(Commentary)

Cloned (Comment) Organism
recombinant expression of wild-type and mutant RNase H RNH domains (residues 427-560) in Escherichia coli strain Rosetta 2 (DE3) as N-terminally His6-SUMO-fused proteins Human immunodeficiency virus 1

Protein Variants

Protein Variants Comment Organism
E438N site-directed mutagenesis, the mutation disrupts RNH folding, viral infectivity is eliminated by the mutation Human immunodeficiency virus 1
E438N/T477A site-directed mutagenesis, addition of the T477A mutation restores correct folding of the RNH domain despite the presence of the E438N mutation Human immunodeficiency virus 1
F440A site-directed mutagenesis, the mutation disrupts RNH folding, viral infectivity is eliminated by the mutation. Molecular dynamics simulations suggest that the T477A mutation affects the processing site by altering relative orientations of secondary structure elements Human immunodeficiency virus 1
F440A/T477A site-directed mutagenesis, addition of the T477A mutation restores correct folding of the RNH domain despite the presence of the F440A mutation Human immunodeficiency virus 1
T477A site-directed mutagenesis, addition of the T477A mutation restores correct folding of the RNH domain despite the presence of the F440A or E438N mutations Human immunodeficiency virus 1

Organism

Organism UniProt Comment Textmining
Human immunodeficiency virus 1 P03366 HIV-1
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Posttranslational Modification

Posttranslational Modification Comment Organism
proteolytic modification the mature form of reverse transcriptase (RT) is a heterodimer comprising the intact 66-kDa subunit (p66) and a smaller 51-kDa subunit (p51) that is generated by removal of most of the RNase H (RNH) domain from a p66 subunit by proteolytic cleavage between residues 440 and 441 Human immunodeficiency virus 1

Purification (Commentary)

Purification (Comment) Organism
recombinant N-terminally His6-SUMO-fused wild-type and mutant RNase H RNH domains (residues 427-560) from Escherichia coli strain Rosetta 2 (DE3) by nickel affinity chromatography and gel filtration, the N-terminal His6-SUMO fusion is removed by digestion with histidine tagged ubiquitin-like-protein specific protease (ULP1) Human immunodeficiency virus 1

Subunits

Subunits Comment Organism
dimer the mature form of reverse transcriptase (RT) is a heterodimer comprising the intact 66-kDa subunit (p66) and a smaller 51-kDa subunit (p51) that is generated by removal of most of the RNase H (RNH) domain from a p66 subunit by proteolytic cleavage between residues 440 and 441 Human immunodeficiency virus 1
More enzyme domain organization, overview Human immunodeficiency virus 1

Synonyms

Synonyms Comment Organism
ribonuclease H
-
 Human immunodeficiency virus 1
ribonuclease H domain in HIV-1 reverse transcriptase
-
 Human immunodeficiency virus 1

General Information

General Information Comment Organism
additional information predictions of sequence tolerance suggest that phenylalanine and tyrosine are structurally preferred at residues 440 and 441, respectively, which are the P1 and P1' substrate residues known to require bulky side chains for substrate specificity. The processing site residues, which are critical for protease substrate specificity and must be exposed to the solvent for efficient processing, also function to maintain proper RNH folding in the p66/p51 heterodimer. Molecular dynamics simulations of wild-type and mutant RNH enzyme domains Human immunodeficiency virus 1