Literature summary for 2.7.7.85 extracted from

Braun, F.; Thomalla, L.; van der Does, C.; Quax, T.E.F.; Allers, T.; Kaever, V.; Albers, S.V.
Cyclic nucleotides in archaea cyclic di-AMP in the archaeon Haloferax volcanii and its putative role (2019), MicrobiologyOpen, 8, e00829 .

Cloned(Commentary)

Cloned (Comment)	Organism
gene dacZ, DNA and amino acid sequence determination and analysis, phylogenetic tree, the Haloferax volcanii genome encodes one diadenylate cyclase (DAC). The dacZ gene is constantly expressed at different growth phases in rich and selective media. Recombinant overexpression of gene dacZ in HAloferax volcanii strain H26, A double auxotrophic background strain H98 (DELTApyrE2 DELTAhdrB) is also transformed with the dacZ-complementing plasmid [p-dacZ+ hdrB+] and subsequently subjected to identical unselective growth conditions. In these control experiments, several colonies auxotrophic for thymidine and hypoxanthine can be detected. Recombinant expression of His10-tagged DacZ in Escherichia coli strain Rosetta	Haloferax volcanii

Cloned (Comment)

Organism

gene dacZ, DNA and amino acid sequence determination and analysis, phylogenetic tree, the Haloferax volcanii genome encodes one diadenylate cyclase (DAC). The dacZ gene is constantly expressed at different growth phases in rich and selective media. Recombinant overexpression of gene dacZ in HAloferax volcanii strain H26, A double auxotrophic background strain H98 (DELTApyrE2 DELTAhdrB) is also transformed with the dacZ-complementing plasmid [p-dacZ+ hdrB+] and subsequently subjected to identical unselective growth conditions. In these control experiments, several colonies auxotrophic for thymidine and hypoxanthine can be detected. Recombinant expression of His10-tagged DacZ in Escherichia coli strain Rosetta

Haloferax volcanii

Protein Variants

Protein Variants	Comment	Organism
additional information	transformation and construction of deletion mutants in Haloferax volcanii strain H26 based on selection with uracil, gene dacZ knockout and overexpression. Because neither gene deletion nor overexpression of dacZ had been possible, a promotor exchange approach is used to examine the effects of altered intracellular c-di-AMP levels. Two mutant strains are created where the native promotor of dacZ is replaced with the tryptophan-inducible promotor (p.tnaA) of the tryptophanase gene tnaA (HVO_0789) or, respectively, with a genetically engineered version of p.tnaA with about 50% reduced activity (with regard to p.tnaA) (p.tnaM3). In comparison to the wild type, the p.tnaA-dacZ strain exhibits in general slightly elevated OD600 values in this condition The p.tnaM3-induced mutant has a different phenotype as it exhibits a prolonged lag phase followed by an extended exponential phase that peaked at an increased maximal OD. Changed intracellular c-di-AMP levels causes increased cell volume in medium with low sodium chloride concentration, phenotypes, overview. When the expression of DacZ is reduced, it leads in turn to decreased c-di-AMP concentrations within the two mutant strains which leads to ta hypo-salt phenotype	Haloferax volcanii

Metals/Ions

Metals/Ions	Comment	Organism	Structure
Mn2+	required, specific for	Haloferax volcanii

Natural Substrates/ Products (Substrates)

Natural Substrates	Organism	Comment (Nat. Sub.)	Natural Products	Comment (Nat. Pro.)	Rev.
2 ATP	Haloferax volcanii	-	2 diphosphate + cyclic di-3',5'-adenylate	-	?
2 ATP	Haloferax volcanii NCIMB 2012	-	2 diphosphate + cyclic di-3',5'-adenylate	-	?
2 ATP	Haloferax volcanii JCM 8879	-	2 diphosphate + cyclic di-3',5'-adenylate	-	?
2 ATP	Haloferax volcanii DS2	-	2 diphosphate + cyclic di-3',5'-adenylate	-	?
2 ATP	Haloferax volcanii DSM 3757	-	2 diphosphate + cyclic di-3',5'-adenylate	-	?
2 ATP	Haloferax volcanii ATCC 29605	-	2 diphosphate + cyclic di-3',5'-adenylate	-	?
2 ATP	Haloferax volcanii NBRC 14742	-	2 diphosphate + cyclic di-3',5'-adenylate	-	?
2 ATP	Haloferax volcanii VKM B-1768	-	2 diphosphate + cyclic di-3',5'-adenylate	-	?

Organism

Organism	UniProt	Comment	Textmining
Haloferax volcanii	D4GZM5	Halobacterium volcanii	-
Haloferax volcanii ATCC 29605	D4GZM5	Halobacterium volcanii	-
Haloferax volcanii DS2	D4GZM5	Halobacterium volcanii	-
Haloferax volcanii DSM 3757	D4GZM5	Halobacterium volcanii	-
Haloferax volcanii JCM 8879	D4GZM5	Halobacterium volcanii	-
Haloferax volcanii NBRC 14742	D4GZM5	Halobacterium volcanii	-
Haloferax volcanii NCIMB 2012	D4GZM5	Halobacterium volcanii	-
Haloferax volcanii VKM B-1768	D4GZM5	Halobacterium volcanii	-

Purification (Commentary)

Purification (Comment)	Organism
recombinant His10-tagged DacZ from Escherichia coli strain Rosetta by nickel affinity chromatography and dialysis	Haloferax volcanii

Source Tissue

Source Tissue	Comment	Organism	Textmining
additional information	gene dacZ is constantly expressed at different growth phases in rich and selective media	Haloferax volcanii	-

Substrates and Products (Substrate)

Substrates	Comment Substrates	Organism	Products	Comment (Products)	Rev.
2 ATP	-	Haloferax volcanii	2 diphosphate + cyclic di-3',5'-adenylate	-	?
2 ATP	-	Haloferax volcanii NCIMB 2012	2 diphosphate + cyclic di-3',5'-adenylate	-	?
2 ATP	-	Haloferax volcanii JCM 8879	2 diphosphate + cyclic di-3',5'-adenylate	-	?
2 ATP	-	Haloferax volcanii DS2	2 diphosphate + cyclic di-3',5'-adenylate	-	?
2 ATP	-	Haloferax volcanii DSM 3757	2 diphosphate + cyclic di-3',5'-adenylate	-	?
2 ATP	-	Haloferax volcanii ATCC 29605	2 diphosphate + cyclic di-3',5'-adenylate	-	?
2 ATP	-	Haloferax volcanii NBRC 14742	2 diphosphate + cyclic di-3',5'-adenylate	-	?
2 ATP	-	Haloferax volcanii VKM B-1768	2 diphosphate + cyclic di-3',5'-adenylate	-	?
additional information	thin layer chromatography product determination. The DAC activity is specific for ATP as substrate and manganese (II) ions as cofactor	Haloferax volcanii	?	-	-
additional information	thin layer chromatography product determination. The DAC activity is specific for ATP as substrate and manganese (II) ions as cofactor	Haloferax volcanii NCIMB 2012	?	-	-
additional information	thin layer chromatography product determination. The DAC activity is specific for ATP as substrate and manganese (II) ions as cofactor	Haloferax volcanii JCM 8879	?	-	-
additional information	thin layer chromatography product determination. The DAC activity is specific for ATP as substrate and manganese (II) ions as cofactor	Haloferax volcanii DS2	?	-	-
additional information	thin layer chromatography product determination. The DAC activity is specific for ATP as substrate and manganese (II) ions as cofactor	Haloferax volcanii DSM 3757	?	-	-
additional information	thin layer chromatography product determination. The DAC activity is specific for ATP as substrate and manganese (II) ions as cofactor	Haloferax volcanii ATCC 29605	?	-	-
additional information	thin layer chromatography product determination. The DAC activity is specific for ATP as substrate and manganese (II) ions as cofactor	Haloferax volcanii NBRC 14742	?	-	-
additional information	thin layer chromatography product determination. The DAC activity is specific for ATP as substrate and manganese (II) ions as cofactor	Haloferax volcanii VKM B-1768	?	-	-

Synonyms

Synonyms	Comment	Organism
Dac	-	Haloferax volcanii
dacZ	-	Haloferax volcanii

Temperature Optimum [°C]

Temperature Optimum [°C]	Temperature Optimum Maximum [°C]	Comment	Organism
37	-	assay at	Haloferax volcanii

pH Optimum

pH Optimum Minimum	pH Optimum Maximum	Comment	Organism
7.5	-	assay at	Haloferax volcanii

General Information

General Information	Comment	Organism
evolution	enzyme DacZ belongs to the proposed DacZ subfamily, sequence and structure comparisons with other DACs. The adenylate cyclase domains of proteins of the DacZ and DacY/CdaZ classes are found in a similar branch of the tree, which is distinct from other bacterial and archaeal classes, and this branch also contains several bacterial proteins. Homologues are identified in most euryarchaeal species, but no DACs are identified in crenarchaeota	Haloferax volcanii
malfunction	a strain with decreased c-di-AMP levels exhibits an increased cell area in hypo-salt medium, implying impaired osmoregulation. Homologous overexpression of DacZ leads to cell death. Changed intracellular c-di-AMP levels causes increased cell volume in medium with low sodium chloride concentration	Haloferax volcanii
physiological function	the DAC encoding gene (dacZ) is essential, but homologous overexpression of DacZ leads to cell death, suggesting the need for tight regulation of c-di-AMP levels. A central target of c-di-AMP signaling in bacteria is cellular osmohomeostasis and a comparable function in euryarchaeon Haloferax volcanii is suggested. Osmoregulation is likely to be a common function of c-di-AMP in bacteria and archaea	Haloferax volcanii