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Literature summary for 3.1.26.13 extracted from

  • Permanasari, E.D.; Yasukawa, K.; Kanaya, S.
    Enzymatic activities of RNase H domains of HIV-1 reverse transcriptase with substrate binding domains of bacterial RNases H1 and H2 (2015), Mol. Biotechnol., 57, 526-538 .
    View publication on PubMed
Search for more literature for 3.1.26.13

Cloned(Commentary)

Cloned (Comment) Organism
recombinant overexpression of wild-type enzyme and chimeric mutants in Escherichia coli strain MIC2067 (DE3) Human immunodeficiency virus 1

Protein Variants

Protein Variants Comment Organism
additional information the isolated RNase H domain of HIV-1 RT (RNHHIV) is inactive, possibly due to the lack of a substrate binding ability, disorder of a loop containing His539, and increased flexibility. To examine whether the activity of RNHHIV is restored by the attachment of TmaHBD or BstNTD to its N-terminus, two chimeric proteins, TmaHBD-RNHHIV and BstNTD-RNHHIV, are constructed and characterized. Both chimeric proteins bind to RNA/DNA hybrid more strongly than RNHHIV and exhibit enzymatic activity in the presence of Mn2+ ions. They do not exhibit activity or exhibited very weak activity in the presence of Mg2+ ions Human immunodeficiency virus 1

Metals/Ions

Metals/Ions Comment Organism Structure
Mg2+ required Human immunodeficiency virus 1
Mn2+ required Human immunodeficiency virus 1

Organism

Organism UniProt Comment Textmining
Human immunodeficiency virus 1
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 HIV-1
-

Purification (Commentary)

Purification (Comment) Organism
recombinant wild-type enzyme and chimeric mutants from Escherichia coli strain MIC2067 (DE3) by dialysis of cell-free enzyme extract, anion and cation exchange chromatography, dialysis, and gel filtration Human immunodeficiency virus 1

Substrates and Products (Substrate)

Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
additional information RNase H activity is determined using oligomeric substrates. 5'-Fluorescein-labeled 29 bp RNA/DNA hybrid (R29/D29), 12 bp RNA/RNA duplex (R12/R12), and 12 bp DNA/DNA duplex (D12/D12) are prepared by hybridizing 5'-fluorescein-labeled 29 b RNA (5'-aauagagaaaaagaaaaaagauggcaaag-3'), 12 b RNA (5'-cggagaugacgg-3'), and 12 b DNA (5'-CGGAGAUGACGG-3') with a 1.5 molar equivalent of the complementary DNA Human immunodeficiency virus 1 ?
-
 ?

Synonyms

Synonyms Comment Organism
ribonuclease H
-
 Human immunodeficiency virus 1
RNase H
-
 Human immunodeficiency virus 1
RNase H domain of HIV-1 reverse transcriptase
-
 Human immunodeficiency virus 1
RNHHIV
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 Human immunodeficiency virus 1

Temperature Optimum [°C]

Temperature Optimum [°C] Temperature Optimum Maximum [°C] Comment Organism
30
-
 assay at Human immunodeficiency virus 1

pH Optimum

pH Optimum Minimum pH Optimum Maximum Comment Organism
8
-
 assay at Human immunodeficiency virus 1

General Information

General Information Comment Organism
additional information the isolated RNase H domain of HIV-1 RT (RNHHIV) is inactive, possibly due to the lack of a substrate binding ability, disorder of a loop containing His539, and increased flexibility. The N-terminal substrate binding domain, termed hybrid binding domain (TmaHBD), from Thermotoga maritima RNase H1 and the N-terminal domain (BstNTD) from Bacillus stearothermophilus RNase H2 both function as an RNA/DNA hybrid binding tag, and greatly increase the substrate binding affinity and Mn2+-dependent activity of RNHHIV but do not restore the Mg2+-dependent activity of RNHHIV Human immunodeficiency virus 1
physiological function ribonuclease H (RNase H) is an enzyme that cleaves RNA strand of RNA/DNA hybrid to produce 5'-phosphate and 3'-hydroxyl termini with a two-metal-ion catalysis mechanism. HIV-1 reverse transcriptase (RT) is a heterodimer consisting of a P66 subunit and a P51 subunit. The P66 subunit contains a C-terminal RNase H domain, which exhibits RNase H activity either in the presence of Mg2+ or Mn2+ ions Human immunodeficiency virus 1