A New Allele, DNASE1*6, of Human Deoxyribonuclease I Polymorphism Encodes an Arg to Cys Substitution Responsible for Its Instability

Yasuda, Toshihiro; Takeshita, Hitoshi; Iida, Reiko; Kogure, Shinichi; Kishi, Koichiro

doi:10.1006/bbrc.1999.0900

Cited by 49 publications

(31 citation statements)

References 15 publications

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“…The DNase I protein has two disulfide bonds, Cys123-Cys126 and Cys195-Cys231, the latter of which has been shown to contribute to the stability of the enzyme [32]. Baron 195 (194) or Cys 231, inducing structural instability through loss of the essential disulfide bond, thereby drastically reducing or abolishing the activity [13,34]. In contrast, the amino acid substituted DNase 1L3 corresponding to p.Tyr261Cys showed an activity level similar to that of the wild-type enzyme.…”

Section: Discussionmentioning

confidence: 99%

Evaluation of all non‐synonymous single nucleotide polymorphisms (SNPs) in the genes encoding human deoxyribonuclease I and I‐like 3 as a functional SNP potentially implicated in autoimmunity

et al. 2013

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The objectives of this study were to evaluate all the non-synonymous single nucleotide polymorphisms (SNPs) in the DNase I and DNase I-like 3 (1L3) genes potentially implicated in autoimmune diseases as a functional SNP in terms of alteration of the activity levels. We examined the genotype distributions of the 32 and 20 non-synonymous SNPs in DNASE1 and DNASE1L3, respectively, in three ethnic groups, and the effect of these SNPs on the DNase activities. Among a total of 44 and 25 SNPs including those characterized in our previous studies [Yasuda et al., Int J Biochem Cell Biol 42 (2010) 1216-1225; Ueki et al. Electrophoresis 32 (2012) 1465-1472], only four and one, respectively, exhibited genetic heterozygosity in one or all of the ethnic groups examined. On the basis of alterations in the activity levels resulting from the corresponding amino acid substitutions, 11 activity-abolishing and 11 activity-reducing SNPs in DNASE1 and two activity-abolishing and five activity-reducing SNPs in DNASE1L3 were confirmed as a functional SNP. Phylogenetic analysis showed that all of the amino acid residues in activity-abolishing SNPs were completely or well conserved in animal DNase I and 1L3 proteins. Although almost all non-synonymous SNPs in both genes that affected the catalytic activity showed extremely low genetic heterogeneity, it seems plausible that a minor allele of 13 activity-abolishing SNPs producing a loss-of-function variant in both the DNase genes would be a direct genetic risk factor for autoimmune diseases. These findings may have clinical implications in relation to the prevalence of autoimmune diseases.

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Section: Discussionmentioning

confidence: 99%

Evaluation of all non‐synonymous single nucleotide polymorphisms (SNPs) in the genes encoding human deoxyribonuclease I and I‐like 3 as a functional SNP potentially implicated in autoimmunity

et al. 2013

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“…However, a useful combination of expression and purification of the recombinant DNase I as an immunogen has so far not been developed. Our previous study indicated that COS-7 cells transiently transfected with expression vector carrying human DNase I cDNA secreted the enzyme in substantial amounts, enabling easy purification of the recombinant enzyme from the culture medium [4]. In this communication, we describe the combined procedure of expression and purification of recombinant DNase I suited for preparation of the enzyme as an immunogen.…”

Section: Introductionmentioning

confidence: 99%

“…Recently, DNase I was suggested to be responsible for the removal of DNA from nuclear antigens at sites of high cell turnover and thus for the prevention of systemic lupus erythematosus [2]. It has been demonstrated by isoelectric focusing analysis that human DNase I exhibits genetic polymorphism, which is controlled by at least 6 alleles on chromosome 16p13.3 [3,4]. In this context, in order to elucidate the intrinsic intra-and extracellular functions of DNase I, as well as its molecular multiplicity, immunological investigations are essential.…”

Section: Introductionmentioning

confidence: 99%

Use of Human Recombinant DNase I Expressed in COS-7 Cells as an Immunogen to Produce a Specific Anti-DNase I Antibody

Takeshita

Yasuda²,

Nakazato

et al. 2001

Exp Clin Immunogenet

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To obtain human deoxyribonuclease I (DNase I) as an immunogen, we have developed a procedure that is more useful and effective than the conventional procedure, which uses human urine as a starting material. In the new procedure, we culture COS-7 cells transfected with expression vector carrying human DNase I cDNA, and then purify the enzyme from the culture medium. The enzyme can be easily isolated to apparent homogeneity by passage through only three chromatography columns. The rabbit antiserum that we used against the recombinant DNase I was not inferior to that used against DNase I from human urine, in terms of both its ability to discriminate DNase I phenotypes and its ability to neutralize enzyme activity. Therefore, our procedure may be useful for producing an antibody specific for human DNase I.

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“…Although DNase I had previously been considered to be merely a digestive enzyme, the presence of its activity in mammalian tissues other than the digestive organs has suggested that it might have other functions in i o [3,4]. With the use of isoelectric focusing, human DNase I has been shown to exhibit genetic polymorphism controlled by at least six alleles [5,6], of which the DNASE1*2 allele might be a risk factor for gastric carcinoma [7]. Mammalian DNases I are classified into three types [pancreatic, parotid and parotid-pancreatic (mixed)], according to their main source, and their sensitivities to low pH are dependent on the type [4].…”

Section: Introductionmentioning

confidence: 99%

Amphibian DNases I are characterized by a C-terminal end with a unique, cysteine-rich stretch and by the insertion of a serine residue into the Ca2+-binding site

Takeshita

Yasuda

Iida

et al. 2001

Biochemical Journal

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We purified four amphibian deoxyribonucleases I from the pancreases of one toad, two frog and one newt species, by using three different column chromatography methods in sequence. Each of the purified enzymes had a molecular mass of approx. 40 kDa and an optimal pH for activity of approx. 8.0. These values were significantly greater than those for other vertebrate DNases I. The full-length cDNA encoding each amphibian DNase I was constructed from the total RNA of the pancreas by using rapid amplification of cDNA ends. Nucleotide sequence analyses revealed two structural characteristics unique to amphibian DNases I : a stretch of approx. 70 amino acids with a high cysteine content (approx. 15 %) in the C-terminal region, and the insertion of a serine residue at position 205 (in a domain containing an essential Ca# + -binding site). Expression analysis of

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A New Allele, DNASE1*6, of Human Deoxyribonuclease I Polymorphism Encodes an Arg to Cys Substitution Responsible for Its Instability

Cited by 49 publications

References 15 publications

Evaluation of all non‐synonymous single nucleotide polymorphisms (SNPs) in the genes encoding human deoxyribonuclease I and I‐like 3 as a functional SNP potentially implicated in autoimmunity

Evaluation of all non‐synonymous single nucleotide polymorphisms (SNPs) in the genes encoding human deoxyribonuclease I and I‐like 3 as a functional SNP potentially implicated in autoimmunity

Use of Human Recombinant DNase I Expressed in COS-7 Cells as an Immunogen to Produce a Specific Anti-DNase I Antibody

Amphibian DNases I are characterized by a C-terminal end with a unique, cysteine-rich stretch and by the insertion of a serine residue into the Ca2+-binding site

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