Literature summary for 3.1.26.13 extracted from

Figiel, M.; Krepl, M.; Poznanski, J.; Golab, A.; Sponer, J.; Nowotny, M.
Coordination between the polymerase and RNase H activity of HIV-1 reverse transcriptase (2017), Nucleic Acids Res., 45, 3341-3352 .

Search for more literature for 3.1.26.13

Cloned(Commentary)

Cloned (Comment)	Organism
recombinant expression of both enzyme subunits, N-terminally His-tagged p66 and p51 subunit mutants C282S of HIV-1, in Escherichia coli strain BL21 Rosetta	Human immunodeficiency virus 1

Protein Variants

Protein Variants	Comment	Organism
C282S	site-directed mutagenesis, the mutation avoids non-specific cross-linking in both subunits, p66 and p51 subunits of HIV-1	Human immunodeficiency virus 1

Inhibitors

Inhibitors	Comment	Organism	Structure
heparin	inhibits substrate binding by the enzyme, in the samples that are pretreated with DTT, the addition of heparin strongly inhibits the reaction and completely eliminates off-register cleavage	Human immunodeficiency virus 1
NaCl	inhibits substrate binding by the enzyme, in the samples that are pretreated with DTT, the addition of salt strongly inhibits the reaction and completely eliminates off-register cleavage	Human immunodeficiency virus 1

Metals/Ions

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

Organism

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

Purification (Commentary)

Purification (Comment)	Organism
recombinant N-terminally His-tagged p66 and p51 subunit mutants C282S from Escherichia coli strain BL21 Rosetta by nickel affinity chromatography, dialysis, heparin affinity chromatography, and gel filtration	Human immunodeficiency virus 1

Substrates and Products (Substrate)

Substrates	Comment Substrates	Organism	Products	Comment (Products)	Rev.	Reac.
additional information	when the RNA/DNA hybrid is immobilized at the polymerase active site, RNase H cleavage occurs, experimentally verifying that the substrate can simultaneously interact with both active sites, analysis of the mechanism of the coordination of the two activities. Cross-linking of HIV-1 RT with RNA/DNA hybrids, inhibition of complex formation by salt and heparin. Purified HIV-1 RT D186C, M184C and Q258C variants are tethered to appropriately modified CL3 substrates, and RNase H activity within the cross-linked complexes is determined, overview	Human immunodeficiency virus 1	?	-	?

Synonyms

Synonyms	Comment	Organism
RNase H	-	Human immunodeficiency virus 1
RNase H activity of HIV-1 reverse transcriptase	-	Human immunodeficiency virus 1

Temperature Optimum [°C]

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

pH Optimum

pH Optimum Minimum	pH Optimum Maximum	Comment	Organism
7.4	-	enzyme domain-substrate cross-linking assay at	Human immunodeficiency virus 1
8	-	RNase H assay at	Human immunodeficiency virus 1

General Information

General Information	Comment	Organism
additional information	molecular dynamics simulations and determination of the conformation of the complex in which the unwound RNA/DNA substrate simultaneously interacts with the polymerase and RNase H active sites. Existence of a transient conformation of the HIV-1 by reverse transcriptase (RT) substrate complex, which is important for modulating and coordinating the enzymatic activities of HIV-1 RT	Human immunodeficiency virus 1
physiological function	replication of human immunodeficiency virus 1 (HIV-1) involves conversion of its single-stranded RNA genome to double-stranded DNA, which is integrated into the genome of the host. This conversion is catalyzed by reverse transcriptase (RT), which possesses DNA polymerase and RNase H domains. When the RNA/DNA hybrid is immobilized at the polymerase active site, RNase H cleavage occurs, experimentally verifying that the substrate can simultaneously interact with both active sites	Human immunodeficiency virus 1

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Release 2023.1 (January 2023)