+ Site Statistics
References:
54,258,434
Abstracts:
29,560,870
PMIDs:
28,072,757
+ Search Articles
+ Subscribe to Site Feeds
Most Shared
PDF Full Text
+ PDF Full Text
Request PDF Full Text
+ Follow Us
Follow on Facebook
Follow on Twitter
Follow on LinkedIn
+ Translate
+ Recently Requested

Topoisomerase I from chicken erythrocytes: purification, characterization, and detection by a deoxyribonucleic acid binding assay



Topoisomerase I from chicken erythrocytes: purification, characterization, and detection by a deoxyribonucleic acid binding assay



Biochemistry 22(8): 2025-2031



We have purified a deoxyribonucleic acid topoisomerase to near homogeneity from the nuclei of mature chicken erythrocytes. The enzyme relaxes supercoiled DNA in the absence of ATP or Mg2+. It is unable to resolve topologically knotted circular duplex DNA. These properties resemble those of type I eukaryotic topoisomerases capable of breaking and rejoining one strand of duplex DNA at a time. The sedimentation value of the protein is 4.4 S. The molecular weight of the reduced, denatured protein is 100K. After elution from sodium dodecyl sulfate (NaDodSO4) gels and renaturation, topoisomerase activity is found in the band at 100K and in minor bands at 95K, 78K, and 73K. The minor bands are likely to be proteolytic fragments since the Mr 100K protein is cleaved by trypsin to fragments of similar or even smaller size with retention of activity. At KCl concentrations suboptimal for the 100K form, the trypsin cleaved form is severalfold more active than the 100K form. Single-stranded DNA, but not duplex DNA or RNA, inhibits DNA relaxing activity, presumably by forming a covalent complex at the enzyme active site. Preincubation of the enzyme with single-stranded DNA leads to the depletion, in NaDodSO4-polyacrylamide gels, of protein bands corresponding to the 100K topoisomerase, its putative proteolytic fragments, and its tryptic fragments. The reaction which leads to band depletion requires active topoisomerase and conditions where single-stranded DNA inhibits relaxing activity. The band depletion technique provides a convenient assay for the polynucleotide binding activity of topoisomerases and possibly other proteins. The function of the enzyme in the inactive nuclei of mature chicken erythrocytes is unclear. The estimated content of chicken erythrocyte topoisomerase per unit DNA is comparable to that in nuclei active in replication and transcription.

(PDF emailed within 1 workday: $29.90)

Accession: 018187206

Download citation: RISBibTeXText

PMID: 6303401


Related references

Deoxyribonucleic acid topoisomerase I from chicken erythrocytes: purification, characterization, and detection by a Dna binding assay. Biochemistry 22(8): 2025-2031, 1983

Purification and properties of type 1 topoisomerase from chicken erythrocytes: mechanism of eukaryotic topoisomerase action. Biochemistry 21(6): 1155-1161, 1982

Total protein, deoxyribonucleic acid, ribonucleic acid, and histone content of isolated nuclei of chicken erythrocytes. Poultry Science 53(6): 2163-2167, 1974

Deoxyribonucleic acid-binding properties of the deoxyribonucleic acid-dependent deoxyribonucleic acid polymerase induced in baby-hamster kidney cells after infection with herpes simplex virus type I. Biochemical Society Transactions 7(5): 1093-1094, 1979

Deoxyribonucleic Acid-Binding Properties of the Deoxyribonucleic Acid-Dependent Deoxyribonucleic Acid Polymerase Induced in Baby-Hamster Kidney Cells after Infection with Herpes Simplex Virus Type I. Biochemical Society Transactions 7(5): 1093-1095, 1979

Deoxyribonucleic acid-deoxyribonucleic acid hybridization assay for replication origin deoxyribonucleic acid of Escherichia coli. Journal of Bacteriology 110(3): 917-925, 1972

Purification and characterization of the 1q subcomponent of canine complement and its use in the 125I-C1q binding assay for detection of immune complexes. American Journal of Veterinary Research 49(6): 865-869, 1988

Ultraviolet light induced preferential cross-linking of histone H3 to deoxyribonucleic acid in chromatin and nuclei of chicken erythrocytes. Biochemistry 21(14): 3419-3427, 1982

Characterization of mitochondrial deoxyribonucleic acid from a series of petite yeast strains by deoxyribonucleic acid-deoxyribonucleic acid hybridization. Biochemistry 13(6): 1067-1075, 1974

Serological Detection of Deoxyribonucleic Acid (DNA) Adsorbed to Formalinized Erythrocytes. Experimental Biology and Medicine 98(2): 300-303, 1958

Purification and characterization of ferro chelatase ec 4.99.1.1 from chicken erythrocytes. Federation Proceedings 41(4): ABSTRACT 5482, 1982

Qualitative and quantitative detection of chicken deoxyribonucleic acid (DNA) in dry dog foods. Journal of Animal Physiology and Animal Nutrition 102 Suppl 1: 37-42, 2018

Qualitative and quantitative detection of chicken deoxyribonucleic acid (DNA) in dry dog foods. Journal of Animal Physiology and Animal Nutrition 102: 37-42, 2018

Characterization of a chicken luteinizing hormone receptor (cLH-R) complementary deoxyribonucleic acid, and expression of cLH-R messenger ribonucleic acid in the ovary. Biology of Reproduction 55(2): 304-309, 1996

The Enzymatic Methylation Of Ribonucleic Acid And Deoxyribonucleic Acid. V. Purification And Properties Of The Deoxyribonucleic Acid-Methylating Activity Of Escherichia Coli. Journal of Biological Chemistry 239: 3858-3865, 1964