Literature summary for 3.1.31.1 extracted from

Kiedrowski, M.R.; Crosby, H.A.; Hernandez, F.J.; Malone, C.L.; McNamara, J.O.; Horswill, A.R.
Staphylococcus aureus Nuc2 is a functional, surface-attached extracellular nuclease (2014), PLoS ONE, 9, e95574.

Cloned(Commentary)

Cloned (Comment)	Organism
gene nuc1, sequence comparison, recombinant expression of C-terminally His6-tagged isozyme Nuc1 in Escherichia coli strain strain ER2566	Staphylococcus aureus
gene nuc2, sequence comparison, the portion of nuc2 encoding amino acids N26-K177 is amplified from AH1263 genomic DNA, recombinant expression of the truncated version as C-terminally His6-tagged isozyme Nuc2, and expression of isozyme Nuc2 as fusion proteins Nuc2-GFP and Nuc2-PhoA, respectively, in Escherichia coli strain strain ER2566, creating strain AH2591. Expression of recombinant chimeric His-tagged fusion proteins in Staphylococcus aureus nuc-deficient mutant strain AH1680	Staphylococcus aureus

Cloned (Comment)

Organism

gene nuc1, sequence comparison, recombinant expression of C-terminally His6-tagged isozyme Nuc1 in Escherichia coli strain strain ER2566

Staphylococcus aureus

gene nuc2, sequence comparison, the portion of nuc2 encoding amino acids N26-K177 is amplified from AH1263 genomic DNA, recombinant expression of the truncated version as C-terminally His6-tagged isozyme Nuc2, and expression of isozyme Nuc2 as fusion proteins Nuc2-GFP and Nuc2-PhoA, respectively, in Escherichia coli strain strain ER2566, creating strain AH2591. Expression of recombinant chimeric His-tagged fusion proteins in Staphylococcus aureus nuc-deficient mutant strain AH1680

Staphylococcus aureus

Protein Variants

Protein Variants	Comment	Organism
additional information	construction of nuc1 and nuc1/nuc2 deletion mutant strains	Staphylococcus aureus
additional information	four chimeric His-tagged fusion proteins are constructed by splicing together: 1. the N-terminal Nuc secretion signal or NucB leader to the Nuc2 C-terminal active domain, or 2. the N-terminal Nuc2 membrane anchor to the NucA and NucB C-terminal active domains, construction of nuc2 and nuc1/nuc2 deletion mutant strains	Staphylococcus aureus

Localization

Localization	Comment	Organism	GeneOntology No.	Textmining
plasma membrane	the isozyme Nuc2 is membrane-bound with the C-terminus facing the extracellular environment, indicating it is a signal-anchored Type II membrane protein	Staphylococcus aureus	5886	-

Metals/Ions

Metals/Ions	Comment	Organism	Structure
additional information	isozyme Nuc2 activity is cation-dependent	Staphylococcus aureus

Organism

Organism	UniProt	Comment	Textmining
Staphylococcus aureus	A0A0H3JPX6	gene nuc2; different USA300 strains, SF8300 and LAC, RN4220, MW2, TCH1516, COL and UAMS-1, gene nuc2	-
Staphylococcus aureus	Q7A6P2	gene nuc1; different USA300 strains, SF8300 and LAC, RN4220, MW2, TCH1516, COL and UAMS-1, gene nuc1	-
Staphylococcus aureus N315	A0A0H3JPX6	gene nuc2; different USA300 strains, SF8300 and LAC, RN4220, MW2, TCH1516, COL and UAMS-1, gene nuc2	-
Staphylococcus aureus N315	Q7A6P2	gene nuc1; different USA300 strains, SF8300 and LAC, RN4220, MW2, TCH1516, COL and UAMS-1, gene nuc1	-

Purification (Commentary)

Purification (Comment)	Organism
recombinant His-tagged truncated isozyme Nuc2 lacking the N-terminal membrane anchor by nickel-affinity chromatography	Staphylococcus aureus

Substrates and Products (Substrate)

Substrates	Comment Substrates	Organism	Products	Comment (Products)	Rev.	Reac.
additional information	the wild-type and truncated mutant isozyme Nuc2 degrades several nucleotide substrates, including Staphylococcus aureus genomic DNA, eukaryotic salmon sperm DNA, double-stranded plasmid DNA, and single-stranded DNA	Staphylococcus aureus	?	-	?
additional information	the wild-type and truncated mutant isozyme Nuc2 degrades several nucleotide substrates, including Staphylococcus aureus genomic DNA, eukaryotic salmon sperm DNA, double-stranded plasmid DNA, and single-stranded DNA	Staphylococcus aureus N315	?	-	?

Synonyms

Synonyms	Comment	Organism
Nuc	-	Staphylococcus aureus
Nuc2	-	Staphylococcus aureus

Temperature Stability [°C]

Temperature Stability Minimum [°C]	Temperature Stability Maximum [°C]	Comment	Organism
75	-	1 h, heat-treated isozyme Nuc1 is still able to degrade the DNA substrate	Staphylococcus aureus
75	-	1 h, heat-treated isozyme Nuc2 is still able to degrade the DNA substrate	Staphylococcus aureus

General Information

General Information	Comment	Organism
additional information	sequence and structure comparisons of isozymes Nuc1 and Nuc2, overview	Staphylococcus aureus
physiological function	isozyme Nuc2 is detrimental to Staphylococcus aureus biofilms, purified isozyme Nuc2 prevents biofilm formation in vitro and modestly decreases biomass in dispersal experiments	Staphylococcus aureus