Human topoisomerase IIα:: Targeting to subchromosomal sites of activity during interphase and mitosis

被引:32
作者
Agostinho, M
Rino, J
Braga, J
Ferreira, F
Steffensen, S
Ferreira, J [1 ]
机构
[1] Univ Lisbon, Fac Med, Inst Histol, P-1649028 Lisbon, Portugal
[2] CIISA, Fac Vet Med, Dept Morphol & Funct, P-1300477 Lisbon, Portugal
关键词
D O I
10.1091/mbc.E03-08-0558
中图分类号
Q2 [细胞生物学];
学科分类号
071009 ; 090102 ;
摘要
Mammalian topoisomerase IIalpha (topo IIalpha) plays a vital role in the removal of topological complexities left on DNA during S phase. Here, we developed a new assay to selectively identify sites of catalytic activity of topo IIalpha with subcellular resolution. We show that topo IIalpha activity concentrates at replicating heterochromatin in late S in a replication-dependent manner and at centric heterochromatin during G2 and M phases. Inhibitor studies indicate that this cell cycle-dependent concentration over heterochromatin is sensitive to chromatin structure. We further show that catalytically active topo IIalpha concentrates along the longitudinal axis of mitotic chromosomes. Finally, we found that catalytically inert forms of the enzyme localize predominantly to splicing speckles in a dynamic manner and that this pool is differentially sensitive to changes in the activities of topo IIalpha itself and RNA polymerase II. Together, our data implicate several previously unsuspected activities in the partitioning of the enzyme between sites of activity and putative depots.
引用
收藏
页码:2388 / 2400
页数:13
相关论文
共 41 条
[1]   Catalytic inhibitors of DNA topoisomerase II [J].
Andoh, T ;
Ishida, R .
BIOCHIMICA ET BIOPHYSICA ACTA-GENE STRUCTURE AND EXPRESSION, 1998, 1400 (1-3) :155-171
[2]   Chromosomes with two intact axial cores are induced by G(2) checkpoint override: Evidence that DNA decatenation is not required to template the chromosome structure [J].
Andreassen, PR ;
Lacroix, FB ;
Margolis, RL .
JOURNAL OF CELL BIOLOGY, 1997, 136 (01) :29-43
[3]  
Austin CA, 1998, BIOESSAYS, V20, P215, DOI 10.1002/(SICI)1521-1878(199803)20:3<215::AID-BIES5>3.0.CO
[4]  
2-Q
[5]   INSITU LOCALIZATION OF DNA TOPOISOMERASE-II, A MAJOR POLYPEPTIDE COMPONENT OF THE DROSOPHILA NUCLEAR MATRIX-FRACTION [J].
BERRIOS, M ;
OSHEROFF, N ;
FISHER, PA .
PROCEEDINGS OF THE NATIONAL ACADEMY OF SCIENCES OF THE UNITED STATES OF AMERICA, 1985, 82 (12) :4142-4146
[6]   Mechanism of action of eukaryotic topoisomerase II and drugs targeted to the enzyme [J].
Burden, DA ;
Osheroff, N .
BIOCHIMICA ET BIOPHYSICA ACTA-GENE STRUCTURE AND EXPRESSION, 1998, 1400 (1-3) :139-154
[7]   The contribution of nuclear compartmentalization to gene regulation [J].
Carmo-Fonseca, M .
CELL, 2002, 108 (04) :513-521
[8]   Dynamics of human DNA topoisomerases IIα and IIβ in living cells [J].
Christensen, MO ;
Larsen, MK ;
Barthelmes, HU ;
Hock, R ;
Andersen, CL ;
Kjeldsen, E ;
Knudsen, BR ;
Westergaard, O ;
Boege, F ;
Mielke, C .
JOURNAL OF CELL BIOLOGY, 2002, 157 (01) :31-44
[9]   A TOPOISOMERASE II-DEPENDENT G2 CYCLE CHECKPOINT IN MAMMALIAN-CELLS [J].
DOWNES, CS ;
CLARKE, DJ ;
MULLINGER, M ;
GIMENEZABIAN, JF ;
CREIGHTON, AM ;
JOHNSON, RT .
NATURE, 1994, 372 (6505) :467-470
[10]   Functional architecture in the cell nucleus [J].
Dundr, M ;
Misteli, T .
BIOCHEMICAL JOURNAL, 2001, 356 (02) :297-310