Analysis of M phase-specific phosphorylation of DNA topoisomerase II

被引:49
作者
Kimura, K [1 ]
Nozaki, N [1 ]
Enomoto, T [1 ]
Tanaka, M [1 ]
Kikuchi, A [1 ]
机构
[1] TOHOKU UNIV,DEPT MOL & CELL BIOL,FAC PHARMACEUT SCI,AOBA KU,SENDAI,MIYAGI 98077,JAPAN
关键词
D O I
10.1074/jbc.271.35.21439
中图分类号
Q5 [生物化学]; Q7 [分子生物学];
学科分类号
071010 ; 081704 ;
摘要
In mammalian cells, two isoforms of DNA topoisomerase II (topo II), topo II alpha and topo II beta, are phosphorylated, The phosphorylation of topo II beta changes its apparent molecular mass determined by SDS-polyacrylamide gel electrophoresis from 180 to 190 kDa in mitotic cells, whereas topo II alpha affects it only slightly (Kimura, K,, Nozaki, N,, Saijo, M,, Kikuchi, A., Ui, M,, and Enomoto, T. (1994) J, Biol, Chem, 269, 24523-24526), Here we examined the stability of the protein and the phosphate moiety of each topo II isoform, as the cells progressed from M to G(1) phase, While its protein moiety remained intact, 75% of the phosphates attached to topo II beta were removed within 4 h after release from mitotic block, On the other hand, 35% of topo II alpha protein and 52% of the attached phosphates disappeared, We verified that M phase-specific phosphorylation had no particular effect on the catalytic activities of both topo II isoforms after extensive phosphatase digestion, We also examined the binding of two isoforms to the nucleus or chromosomes, In logarithmically growing cells, both isoforms were extracted from nuclei at the same concentrations of NaCl, From the mitotic chromosomes, topo II beta was extracted at much lower concentrations of NaCl than topo II alpha.
引用
收藏
页码:21439 / 21445
页数:7
相关论文
共 40 条
[1]  
ACKERMAN P, 1988, J BIOL CHEM, V263, P12653
[2]   CHROMOSOME ASSEMBLY INVITRO - TOPOISOMERASE-II IS REQUIRED FOR CONDENSATION [J].
ADACHI, Y ;
LUKE, M ;
LAEMMLI, UK .
CELL, 1991, 64 (01) :137-148
[3]   CELL-CYCLE-DEPENDENT PHOSPHORYLATION AND ACTIVITY OF CHINESE-HAMSTER OVARY TOPOISOMERASE-II [J].
BURDEN, DA ;
GOLDSMITH, LJ ;
SULLIVAN, DM .
BIOCHEMICAL JOURNAL, 1993, 293 :297-304
[4]   PHOSPHORYLATION OF THE ALPHA-ISOFORM AND BETA-ISOFORM OF DNA TOPOISOMERASE-II IS QUALITATIVELY DIFFERENT IN INTERPHASE AND MITOSIS IN CHINESE-HAMSTER OVARY CELLS [J].
BURDEN, DA ;
SULLIVAN, DM .
BIOCHEMISTRY, 1994, 33 (49) :14651-14655
[5]  
CARDENAS ME, 1993, J CELL SCI, V104, P219
[6]   CASEIN KINASE-II PHOSPHORYLATES THE EUKARYOTE-SPECIFIC C-TERMINAL DOMAIN OF TOPOISOMERASE-II INVIVO [J].
CARDENAS, ME ;
DANG, Q ;
GLOVER, CVC ;
GASSER, SM .
EMBO JOURNAL, 1992, 11 (05) :1785-1796
[7]  
CARON PR, 1993, MOLECULAR BIOLOGY OF DNA TOPOISOMERASES AND ITS APPLICATION CHEMOTHERAPY, P1
[8]   DNA TOPOISOMERASE-II MUTANT OF SACCHAROMYCES-CEREVISIAE - TOPOISOMERASE-II IS REQUIRED FOR SEGREGATION OF DAUGHTER MOLECULES AT THE TERMINATION OF DNA-REPLICATION [J].
DINARDO, S ;
VOELKEL, K ;
STERNGLANZ, R .
PROCEEDINGS OF THE NATIONAL ACADEMY OF SCIENCES OF THE UNITED STATES OF AMERICA-BIOLOGICAL SCIENCES, 1984, 81 (09) :2616-2620
[9]   BIOCHEMICAL AND PHARMACOLOGICAL PROPERTIES OF P170 AND P180 FORMS OF TOPOISOMERASE-II [J].
DRAKE, FH ;
HOFMANN, GA ;
BARTUS, HF ;
MATTERN, MR ;
CROOKE, ST ;
MIRABELLI, CK .
BIOCHEMISTRY, 1989, 28 (20) :8154-8160
[10]   TOPOISOMERASE-II IS A STRUCTURAL COMPONENT OF MITOTIC CHROMOSOME SCAFFOLDS [J].
EARNSHAW, WC ;
HALLIGAN, B ;
COOKE, CA ;
HECK, MMS ;
LIU, LF .
JOURNAL OF CELL BIOLOGY, 1985, 100 (05) :1706-1715