Literature summary for 3.1.13.2 extracted from

Lim, D.; Gregorio, G.G.; Bingman, C.; Martinez-Hackert, E.; Hendrickson, W.A.; Goff, S.P.
Crystal structure of the moloney murine leukemia virus RNase H domain (2006), J. Virol., 80, 8379-8389.

Cloned(Commentary)

Cloned (Comment)	Organism
expression of wild-type and mutant enzymes, containing a Factor Xa cleavage site and a GST-tag, in Escherichia coli strain BL21(DE3)	Moloney murine leukemia virus

Crystallization (Commentary)

Crystallization (Comment)	Organism
purified recombinant mutant Mo-MLV RNase H lacking the putative helix C complexed with the DNA/RNA hybrid substrate, DELTAC monomers and dimers, equilibration against a reservoir solution of 30% PEG 4000 and 0.2 M ammonium sulfate, protein solution, containing 7-10 mg/ml, 10 mM HEPES, pH 7.0, 150 mM NaCl, 5 mM DTT, and 0.1 mM EDTA, is mixed with reservoir solution at pH 5.2-6.0, DELTAC monomer crystallization fails to yield single, large crystals, DELTAC monomer crystals are soaked in 15% PEG 4000, 20% PEG 400, 0.1 M ammonium sulfate, 150 mM NaCl, 1% 2-propanol, 1.25% PEG MME 550, 5 mM MES, pH 6.5, and 0.5 mM zinc sulfate for 5 min, X-ray diffractions tructure determination and analysis at 1.6 A resolution, modelling	Moloney murine leukemia virus

Crystallization (Comment)

Organism

purified recombinant mutant Mo-MLV RNase H lacking the putative helix C complexed with the DNA/RNA hybrid substrate, DELTAC monomers and dimers, equilibration against a reservoir solution of 30% PEG 4000 and 0.2 M ammonium sulfate, protein solution, containing 7-10 mg/ml, 10 mM HEPES, pH 7.0, 150 mM NaCl, 5 mM DTT, and 0.1 mM EDTA, is mixed with reservoir solution at pH 5.2-6.0, DELTAC monomer crystallization fails to yield single, large crystals, DELTAC monomer crystals are soaked in 15% PEG 4000, 20% PEG 400, 0.1 M ammonium sulfate, 150 mM NaCl, 1% 2-propanol, 1.25% PEG MME 550, 5 mM MES, pH 6.5, and 0.5 mM zinc sulfate for 5 min, X-ray diffractions tructure determination and analysis at 1.6 A resolution, modelling

Moloney murine leukemia virus

Protein Variants

Protein Variants	Comment	Organism
additional information	construction of a mutant Mo-MLV RNase H lacking the putative helix C, surface mapping and substrate binding determinants, overview	Moloney murine leukemia virus

Metals/Ions

Metals/Ions	Comment	Organism	Structure
Mg2+	coordination of the magnesium ion within the catalytic core comprised of the highly conserved acidic residues D524, E562, and D583	Moloney murine leukemia virus

Natural Substrates/ Products (Substrates)

Natural Substrates	Organism	Comment (Nat. Sub.)	Natural Products	Comment (Nat. Pro.)	Rev.	Reac.
RNA/DNA hybrid + H2O	Moloney murine leukemia virus	-	?	-	?

Organism

Organism	UniProt	Comment	Textmining
Moloney murine leukemia virus	-	i.e. Mo-MLV	-

Purification (Commentary)

Purification (Comment)	Organism
recombinant wild-type and mutant enzymes, containing a Factor Xa cleavage site and a GST-tag, from Escherichia coli strain BL21(DE3) by glutathione affinity chromatograpy and by cleavage through Factor Xa, followed by anion and cation exchang chromatography, and gel filtration	Moloney murine leukemia virus

Reaction

Reaction	Comment	Organism	Reaction ID
3'-end directed exonucleolytic cleavage of viral RNA-DNA hybrid	RNase H domain structure and mechanism of catalysis	Moloney murine leukemia virus	a

Substrates and Products (Substrate)

Substrates	Comment Substrates	Organism	Products	Comment (Products)	Rev.	Reac.
RNA/DNA hybrid + H2O	-	Moloney murine leukemia virus	?	-	?
RNA/DNA hybrid + H2O	secondary structure and substrate binding, overview	Moloney murine leukemia virus	?	-	?

Subunits

Subunits	Comment	Organism
More	surface charge mapping of the Mo-MLV structure reveals a high density of basic charges on one side of the enzyme, secondary structure and substrate binding, overview, structure comparisons, overview	Moloney murine leukemia virus

Synonyms

Synonyms	Comment	Organism
RNase H	-	Moloney murine leukemia virus