Abstract
We studied the mutation effect of one of the putative loop residues Thr792 in human DNA topoisomerase IIα (TOP2α). Thr792 mutants were expressed from high or low copy plasmids in a temperature sensitive yeast strain deficient in TOP2 (top2-1). When expressed from a high copy plasmid, mutants with small side chains complemented the yeast defect; however, from a low copy plasmid, only wild-type, Ser, and Cys substitution mutants complemented the yeast defect. Interestingly, at the permissive temperature other mutants (e.g., Val, Gly, and Glu substitutions) showed the dominant negative effect to the top2-1 allele, which was not observed by the control α4-helix mutants. T792E mutant was 10-fold less active than wild-type and the T792P had no decatenation activity in vitro. These results suggest that Thr792 in human TOP2α is involved in enzyme catalysis.
| Original language | English |
|---|---|
| Pages (from-to) | 46-51 |
| Number of pages | 6 |
| Journal | Biochemical and Biophysical Research Communications |
| Volume | 303 |
| Issue number | 1 |
| DOIs | |
| Publication status | Published - 28-03-2003 |
| Externally published | Yes |
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All Science Journal Classification (ASJC) codes
- Biophysics
- Biochemistry
- Molecular Biology
- Cell Biology
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Function of the loop residue Thr792 in human DNA topoisomerase IIα. / Suda, Namiko; Nakagawa, Yoshiyuki; Kikuchi, Akihiko; Sawada, Masako; Takami, Yuko; Funahashi, Hiro Omi; Nakao, Akimasa; Yoshida, Shonen; Suzuki, Motoshi.
In: Biochemical and Biophysical Research Communications, Vol. 303, No. 1, 28.03.2003, p. 46-51.Research output: Contribution to journal › Article
TY - JOUR
T1 - Function of the loop residue Thr792 in human DNA topoisomerase IIα
AU - Suda, Namiko
AU - Nakagawa, Yoshiyuki
AU - Kikuchi, Akihiko
AU - Sawada, Masako
AU - Takami, Yuko
AU - Funahashi, Hiro Omi
AU - Nakao, Akimasa
AU - Yoshida, Shonen
AU - Suzuki, Motoshi
PY - 2003/3/28
Y1 - 2003/3/28
N2 - We studied the mutation effect of one of the putative loop residues Thr792 in human DNA topoisomerase IIα (TOP2α). Thr792 mutants were expressed from high or low copy plasmids in a temperature sensitive yeast strain deficient in TOP2 (top2-1). When expressed from a high copy plasmid, mutants with small side chains complemented the yeast defect; however, from a low copy plasmid, only wild-type, Ser, and Cys substitution mutants complemented the yeast defect. Interestingly, at the permissive temperature other mutants (e.g., Val, Gly, and Glu substitutions) showed the dominant negative effect to the top2-1 allele, which was not observed by the control α4-helix mutants. T792E mutant was 10-fold less active than wild-type and the T792P had no decatenation activity in vitro. These results suggest that Thr792 in human TOP2α is involved in enzyme catalysis.
AB - We studied the mutation effect of one of the putative loop residues Thr792 in human DNA topoisomerase IIα (TOP2α). Thr792 mutants were expressed from high or low copy plasmids in a temperature sensitive yeast strain deficient in TOP2 (top2-1). When expressed from a high copy plasmid, mutants with small side chains complemented the yeast defect; however, from a low copy plasmid, only wild-type, Ser, and Cys substitution mutants complemented the yeast defect. Interestingly, at the permissive temperature other mutants (e.g., Val, Gly, and Glu substitutions) showed the dominant negative effect to the top2-1 allele, which was not observed by the control α4-helix mutants. T792E mutant was 10-fold less active than wild-type and the T792P had no decatenation activity in vitro. These results suggest that Thr792 in human TOP2α is involved in enzyme catalysis.
UR - http://www.scopus.com/inward/record.url?scp=0242286178&partnerID=8YFLogxK
UR - http://www.scopus.com/inward/citedby.url?scp=0242286178&partnerID=8YFLogxK
U2 - 10.1016/S0006-291X(03)00297-3
DO - 10.1016/S0006-291X(03)00297-3
M3 - Article
C2 - 12646164
AN - SCOPUS:0242286178
VL - 303
SP - 46
EP - 51
JO - Biochemical and Biophysical Research Communications
JF - Biochemical and Biophysical Research Communications
SN - 0006-291X
IS - 1
ER -