DNA topoisomerases I and II in human mature sperm cells: characterization and unique properties.
BACKGROUND
The condensed state of the human sperm's chromatin is essential for the compact structure of the spermatozoon head, which is important for the fertilization process. The enzymes DNA topoisomerases (topo I and topo II) are responsible for the topological structure of the chromatin in somatic cells. The activities and the characterization of topoisomerases in mature human sperm cells have not been previously investigated.


METHODS
Sperm cells were purified from the semen of healthy donors by standard procedures and assays measuring the activities, protein size and sensitivity to inhibitors of topoisomerases were performed.


RESULTS
Topo I and topo II DNA relaxation activities are present in nuclear extracts derived from human sperm. The sperm topo I activity is inhibited by camptothecin, similarly to the somatic enzyme. An 85 kDa sperm protein, compared with the 100 kDa somatic topo IB enzyme, reacted with anti-human topo I antibody. Sperm topo II lacks the DNA decatenation activity of the somatic enzyme and a 97 kDa protein, compared with the 170 kDa somatic topo IIalpha enzyme, was detected with anti-human topo II antibody. Sperm nuclear extracts contained inhibitors of somatic topo II decatenation activity.


CONCLUSIONS
Topoisomerase I and II activities as well as topo I and topo II proteins are present in mature human sperm cells. These enzymes possess unique properties compared with their somatic counterparts.
IVF unit, Department of Obstetrics and Gynaecology, Soroka University Medical Center, Faculty of Health Sciences, Ben-Gurion University, Beer-Sheva 84105, Israel.
I
I
Har-Vardi
R
R
Mali
M
M
Breietman
Y
Y
Sonin
S
S
Albotiano
E
E
Levitas
G
G
Potashnik
E
E
Priel
D002166:Camptothecin / Q000494:pharmacology; D004264:DNA Topoisomerases, Type I / Q000276:immunology / Q000302:isolation & purification / Q000378:metabolism*; D004250:DNA Topoisomerases, Type II / Q000276:immunology / Q000302:isolation & purification / Q000378:metabolism*; D005047:Etoposide / Q000494:pharmacology; D006801:Humans; D008297:Male; D008970:Molecular Weight; D013094:Spermatozoa / Q000201:enzymology*; D059004:Topoisomerase I Inhibitors; D059005:Topoisomerase II Inhibitors
D059004:Topoisomerase I Inhibitors; D059005:Topoisomerase II Inhibitors; D005047:Etoposide; D004264:DNA Topoisomerases, Type I; D004250:DNA Topoisomerases, Type II; D002166:Camptothecin