RRC ID |
68509
|
Author |
Ishida R, Takashima R, Koujin T, Shibata M, Nozaki N, Seto M, Mori H, Haraguchi T, Hiraoka Y.
|
Title |
Mitotic specific phosphorylation of serine-1212 in human DNA topoisomerase IIalpha.
|
Journal |
Cell Struct Funct
|
Abstract |
It is known that topoisomerase IIalpha is phosphorylated by several kinases. To elucidate the role of phosphorylation of topoisomerase IIalpha in the cell cycle, we have examined the cell cycle behavior of phosphorylated topoisomerase IIalpha in HeLa cells using antibodies against several phospho-oligopeptides of this enzyme. Here we demonstrate that serine1212 in topoisomerase IIalpha is phosphorylated only in the mitotic phase. Using an antibody against an oligopeptide containing phosphoserine-1212 in topoisomerase IIalpha (PS1212), subcellular localization of topoisomerase IIalpha phosphorylated at serine1212 was examined by indirect immunofluorescence staining, and compared with that of overall topoisomerase IIalpha. Serine1212-phosphorylated topoisomerase IIalpha was localized specifically on mitotic chromosomes, but not on interphase chromosomes; this result contrasts with overall topoisomerase IIalpha which was observed on chomosomes in both interphase and mitosis. Serine1212-phosphorylated topoisomerase lIalpha first appeared on chromosome arms in prophase, became concentrated on the centromeres in metaphase, and disappeared in early telophase. In addition, ICRF-193, a catalytic inhibitor of topoisomerase II, prevented accumulation of serine1212-phosphorylated topoisomerase IIalpha at the centromeres. These results indicate that serine1212 of topoisomerase IIalpha is phosphorylated specifically during mitosis, and suggest that the serine1212-phosphorylated topoisomerase IIalpha acts on resolving topological constraint progressively from the chromosome arm to the centromere during metaphase chromosome condensation.
|
Volume |
26(4)
|
Pages |
215-26
|
Published |
2001-8-1
|
DOI |
10.1247/csf.26.215
|
PMID |
11699638
|
MeSH |
Antibodies, Monoclonal / immunology
Antigens, Neoplasm
Cell Cycle
Centromere / enzymology
Chromosomes / enzymology
DNA Topoisomerases, Type II / chemistry*
DNA Topoisomerases, Type II / immunology
DNA Topoisomerases, Type II / metabolism*
DNA-Binding Proteins
Diketopiperazines
Enzyme Inhibitors / pharmacology
Flow Cytometry
Fluorescent Antibody Technique, Indirect
HeLa Cells
Humans
Microscopy, Fluorescence
Mitosis
Phosphorylation
Piperazines / pharmacology
Serine / metabolism*
Topoisomerase II Inhibitors
|
IF |
2.429
|
Resource |
Human and Animal Cells |
HeLa(RCB0007) |