Characterization and immunological identification of cDNA clones encoding two human DNA topoisomerase II isozymes

Journal Article · Fri Dec 01 00:00:00 EST 1989 · Proceedings of the National Academy of Sciences of the United States of America; (USA)

DOI:https://doi.org/10.1073/pnas.86.23.9431· OSTI ID:6855365

Chung, T D.Y.; Drake, F H; Tan, K B; Per, S R; Crooke, S T; Mirabelli, C K ^[1]

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1. Smith Kline and French Labs., King of Prussia, PA (USA)

Several DNA topoisomerase II partial cDNA clones obtained from a human Raji-HN2 cDNA library were sequenced and two classes of nucleotide sequences were found. One member of the first class, SP1, was identical to an internal fragment of human HeLa cell Topo II cDNA described earlier. A member of the second class, SP11, shared extensive nucleotide (75%) and predicted peptide (92%) sequence similarities with the first two-thirds of HeLa Topo II. Each class of cDNAs hybridized to unique, nonoverlapping restriction enzyme fragments of genomic DNA from several human cell lines. Synthetic 24-mer oligonucleotide probes specific for each cDNA class hybridized to 6.5-kilobase mRNAs; furthermore, hybridization of probe specific for one class was not blocked by probe specific for the other. Antibodies raised against a synthetic SP1-encoded dodecapeptide specifically recognized the 170-kDa form of Topo II, while antibodies raised against the corresponding SP11-encoded dodecapeptide, or a second unique SP11-encoded tridecapeptide, selectively recognized the 180-kDa form of Topo II. These data provide genetic and immunochemical evidence for two Topo II isozymes.

• OSTI ID: 6855365
• Journal Information: Proceedings of the National Academy of Sciences of the United States of America; (USA), Vol. 86:23; ISSN 0027-8424
• Country of Publication: United States
• Language: English

Related Subjects

59 BASIC BIOLOGICAL SCIENCES
ISOMERASES
ISOENZYMES
RECOMBINANT DNA
DNA SEQUENCING
ANTIBODIES
DNA HYBRIDIZATION
GENES
GENETIC MAPPING
GENETICS
HELA CELLS
IMMUNOLOGY
MAN
OLIGONUCLEOTIDES
PHOSPHORUS 32
ANIMALS
BETA DECAY RADIOISOTOPES
BETA-MINUS DECAY RADIOISOTOPES
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DAYS LIVING RADIOISOTOPES
DNA
ENZYMES
HYBRIDIZATION
ISOTOPES
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MAMMALS
MAPPING
NUCLEI
NUCLEIC ACIDS
ODD-ODD NUCLEI
ORGANIC COMPOUNDS
PHOSPHORUS ISOTOPES
PRIMATES
RADIOISOTOPES
STRUCTURAL CHEMICAL ANALYSIS
VERTEBRATES
550201* - Biochemistry- Tracer Techniques