title: DLAD, a novel mammalian divalent cation-independent endonuclease with homology to DNase II.
abstract: In this report, we describe the molecular cloning and characterization of DLAD, a novel mammalian deoxy-ribonuclease homologous to DNase II. The full length cDNA for mouse DLAD has been cloned by polymerase chain reaction. The cDNA contains a 1065 bp open reading frame (ORF) encoding a 354 amino acid protein with a calculated molecular mass of 40 767. The predicted protein for DLAD shares 34.4% identity with DNase II. DLAD is also homologous to three predicted proteins, C07B5.5, F09G8.2 and K04H4.6, from the nematode Caenorhabditis elegans. Furthermore, the third ORF of the fowlpox virus genome is found to encode a DLAD homologue showing 37. 1% identity at the amino acid level. Northern blot analysis reveals that expression of the DLAD mRNA is highly restricted to the liver. DLAD mainly exists as a cytoplasmic protein with divalent cation-independent endonuclease activity and cleaves DNA to produce 3'-phosphoryl/5'-hydroxyl ends. It is active under a wide range of pH with maximum activity at pH 5.2. Among known DNase inhibitors tested, aurintricarboxylic acid and Zn(2+)are found to be effective inhibitors of the DLAD activity.
authors.affiliation: Department of Biochemistry, Faculty of Pharmaceutical Sciences, Science University of Tokyo, 12 Funagawara-machi, Ichigaya, Shinjuku-ku, Tokyo 162-0826, Japan.
authors.forename: D
authors.initials: D
authors.lastname: Shiokawa
authors.forename: S
authors.initials: S
authors.lastname: Tanuma
mesh_terms: D000595:Amino Acid Sequence; D000818:Animals; D001483:Base Sequence; D002413:Cations, Divalent / Q000378:metabolism; D003001:Cloning, Molecular; D004247:DNA / Q000378:metabolism; D018076:DNA, Complementary / Q000737:chemistry / Q000302:isolation & purification; D004706:Endodeoxyribonucleases / Q000737:chemistry / Q000235:genetics* / Q000378:metabolism*; D020224:Expressed Sequence Tags; D006367:HeLa Cells; D006801:Humans; D006868:Hydrolysis; D051379:Mice; D008969:Molecular Sequence Data
chemical_list: D002413:Cations, Divalent; D018076:DNA, Complementary; D004247:DNA; D004706:Endodeoxyribonucleases; C019329:deoxyribonuclease II