TY - JOUR
T1 - Analysis of functional domain organization in DNA topoisomerase II from humans and Saccharomyces cerevisiae
AU - Jensen, Sanne
AU - Andersen, Anni H.
AU - Kjeldsen, Eigil
AU - Biersack, Harald
AU - Olsen, Eva H.N.
AU - Andersen, Torben B.
AU - Westergaard, Ole
AU - Jakobsen, Bent K.
PY - 1996
Y1 - 1996
N2 - The functional domain structure of human DNA topoisomerase IIα and Saccharomyces cerevisiae DNA topoisomerase 11 was studied by investigating the abilities of insertion and deletion mutant enzymes to support mitotic growth and catalyze transitions in DNA topology in vitro. Alignment of the human topoisomerase IIα and S. cerevisiae topoisomerase II sequences defined 13 conserved regions separated by less conserved or differently spaced sequences. The spatial tolerance of the spacer regions was addressed by insertion of linkers. The importance of the conserved regions was assessed through deletion of individual domains. We found that the exact spacing between most of the conserved domains is noncritical, as insertions in the spacer regions were tolerated with no influence on complementation ability. All conserved domains, however, are essential for sustained mitotic growth of S. cerevisiae and for enzymatic activity in vitro. A series of topoisomerase II carboxy-terminal truncations were investigated with respect to the ability to support viability, cellular localization, and enzymatic properties. The analysis showed that the divergent carboxy-terminal region of human topoisomerase IIα is dispensable for catalytic activity but contains elements that specifically locate the protein to the nucleus.
AB - The functional domain structure of human DNA topoisomerase IIα and Saccharomyces cerevisiae DNA topoisomerase 11 was studied by investigating the abilities of insertion and deletion mutant enzymes to support mitotic growth and catalyze transitions in DNA topology in vitro. Alignment of the human topoisomerase IIα and S. cerevisiae topoisomerase II sequences defined 13 conserved regions separated by less conserved or differently spaced sequences. The spatial tolerance of the spacer regions was addressed by insertion of linkers. The importance of the conserved regions was assessed through deletion of individual domains. We found that the exact spacing between most of the conserved domains is noncritical, as insertions in the spacer regions were tolerated with no influence on complementation ability. All conserved domains, however, are essential for sustained mitotic growth of S. cerevisiae and for enzymatic activity in vitro. A series of topoisomerase II carboxy-terminal truncations were investigated with respect to the ability to support viability, cellular localization, and enzymatic properties. The analysis showed that the divergent carboxy-terminal region of human topoisomerase IIα is dispensable for catalytic activity but contains elements that specifically locate the protein to the nucleus.
UR - http://www.scopus.com/inward/record.url?scp=0029890655&partnerID=8YFLogxK
U2 - 10.1128/MCB.16.7.3866
DO - 10.1128/MCB.16.7.3866
M3 - Journal articles
C2 - 8668204
AN - SCOPUS:0029890655
SN - 0270-7306
VL - 16
SP - 3866
EP - 3877
JO - Molecular and Cellular Biology
JF - Molecular and Cellular Biology
IS - 7
ER -