Molecular cloning of partial cDNAs for rat DNA topoisomerase II isoforms and their differential expression in brain development.
cDNA segments for DNA topoisomerase II were amplified from rat brain RNA after reverse transcription by the polymerase chain reaction, using degenerate oligonucleotide primers deduced from the conserved regions of topoisomerase II of higher eukaryotes. The cDNA product from a successful amplification was homogeneous in length but heterogeneous in sequence. Restriction mapping of the cloned cDNA fragments revealed that they consisted of two distinct sequence groups. DNA sequencing of representative clones from each group, designated A and B, showed that they are highly homologous to cDNAs of human topoisomerase II isoforms, alpha and beta, respectively. Northern blot analysis indicated that the transcript level for rat topoisomerase II alpha was high in embryonic brain and in the cerebellum of 2-day newborns, followed by rapid decrease to a undetectable level at 4 weeks after birth. In contrast, rat topoisomerase II beta transcript was present throughout the embryonic and postnatal stages. In the developing cerebellum, cells expressing topoisomerase II alpha were confirmed exclusively to the outer mitotic zone of the external granular layer, whereas the transcript of topoisomerase II beta was detected over the entire cortical region. These results clearly indicate that the isoform alpha is expressed only in proliferating cells. The differential expression of topoisomerase II isozymes was also observed among developed tissues. Therefore, the isozymes are most likely to be involved in the following different physiological processes: topoisomerase II alpha in cell proliferation, and topoisomerase II beta in some processes unrelated to cell proliferation.
Tsutsui
K
K
Department of Neurology, Okayama University Medical School, Japan.
Tsutsui
K
K
Okada
S
S
Watanabe
M
M
Shohmori
T
T
Seki
S
S
Inoue
Y
Y
D000595:Amino Acid Sequence; D000818:Animals; D001483:Base Sequence; D015139:Blotting, Southern; D001921:Brain / Q000201:enzymology / Q000254:growth & development*; D003001:Cloning, Molecular; D004247:DNA / Q000737:chemistry / Q000235:genetics*; D004250:DNA Topoisomerases, Type II / Q000737:chemistry / Q000235:genetics*; D015870:Gene Expression; D006801:Humans; D017403:In Situ Hybridization; D007527:Isoenzymes / Q000737:chemistry / Q000235:genetics*; D008969:Molecular Sequence Data; D016133:Polymerase Chain Reaction; D012333:RNA, Messenger / Q000032:analysis; D051381:Rats; D017207:Rats, Sprague-Dawley; D015183:Restriction Mapping; D017386:Sequence Homology, Amino Acid; D014018:Tissue Distribution
D007527:Isoenzymes; D012333:RNA, Messenger; D004247:DNA; D004250:DNA Topoisomerases, Type II