Isolation and characterization of mAMSA-hypersensitive mutants. Cytotoxicity of Top2 covalent complexes containing DNA single strand breaks.

The Journal of Biological Chemistry
Anna T Rogojina, John L Nitiss

Abstract

Topoisomerase II (Top2) is the primary target for active anti-cancer agents. We developed an efficient approach for identifying hypersensitive Top2 mutants and isolated a panel of mutants in yeast Top2 conferring hypersensitivity to the intercalator N-[4-(9-acridinylamino)-3-methoxyphenyl]methanesulphonanilide (mAMSA). Some mutants conferred hypersensitivity to etoposide as well as mAMSA, whereas other mutants exhibited hypersensitivity only to mAMSA. Two mutants in Top2, changing Pro(473) to Leu and Gly(737) to Val, conferred extraordinary hypersensitivity to mAMSA and were chosen for further characterization. The mutant proteins were purified, and their biochemical activities were assessed. Both mutants encode enzymes that are hypersensitive to inhibition by mAMSA and other intercalating agents and exhibited elevated levels of mAMSA-induced Top2:DNA covalent complexes. While Gly(737) --> Val Top2p generated elevated levels of Top2-mediated double strand breaks in vitro, the Pro(473) --> Leu mutant protein showed only a modest increase in Top2-mediated double strand breaks but much higher levels of Top2-mediated single strand breaks. In addition, the Pro(473) --> Leu mutant protein also generated high levels of mAMSA-stabilize...Continue Reading

References

Mar 25, 1992·Nucleic Acids Research·D GietzR H Schiestl
Sep 1, 1991·Proceedings of the National Academy of Sciences of the United States of America·B Y BuggD P Suttle
Oct 1, 1988·Proceedings of the National Academy of Sciences of the United States of America·J Nitiss, J C Wang
Nov 8, 1994·Proceedings of the National Academy of Sciences of the United States of America·C H Freudenreich, K N Kreuzer
Jan 1, 1994·Cancer Chemotherapy and Pharmacology·J L Nitiss
Jun 1, 1996·European Journal of Cancer : Official Journal for European Organization for Research and Treatment of Cancer (EORTC) [and] European Association for Cancer Research (EACR)·J L Nitiss, W T Beck
Aug 1, 1998·The Journal of Biological Chemistry·Q Liu, J C Wang
Sep 28, 1998·Biochimica Et Biophysica Acta·J L Nitiss
Sep 28, 1998·Biochimica Et Biophysica Acta·P S Kingma, N Osheroff
Feb 3, 1999·Proceedings of the National Academy of Sciences of the United States of America·Q Liu, J C Wang
Sep 25, 1999·The Journal of Biological Chemistry·D StrumbergY Pommier
Mar 20, 2001·Current Pharmaceutical Design·V E Anderson, N Osheroff
Mar 27, 2001·Annual Review of Pharmacology and Toxicology·T K Li, L F Liu
Jun 4, 2002·Nature Reviews. Molecular Cell Biology·James C Wang
Jul 4, 2002·Cancer Investigation·Jerrylaine V Walker, John L Nitiss
Feb 12, 2004·Eukaryotic Cell·Mobeen Malik, John L Nitiss
Mar 24, 2004·The Journal of Biological Chemistry·Jerrylaine V WalkerJohn L Nitiss
Jan 14, 2005·The Journal of Biological Chemistry·Jerrylaine VaughnJohn L Nitiss
Jun 6, 2006·Proceedings of the National Academy of Sciences of the United States of America·Karin C NitissJohn L Nitiss
Jul 5, 2006·Molecular Cancer Therapeutics·Mobeen MalikJohn L Nitiss

❮ Previous
Next ❯

Citations

Apr 21, 2009·Nature Reviews. Cancer·John L Nitiss
Oct 9, 2012·Biochemical Pharmacology·Brian B HasinoffJack C Yalowich
Oct 20, 2011·The Journal of Toxicological Sciences·Yoko MoritaIsao Kuraoka
Oct 14, 2020·Proceedings of the National Academy of Sciences of the United States of America·Nicole StantialJohn L Nitiss

❮ Previous
Next ❯

Related Concepts

Related Feeds

Allergy and Asthma

Allergy and asthma are inflammatory disorders that are triggered by the activation of an allergen-specific regulatory t cell. These t cells become activated when allergens are recognized by allergen-presenting cells. Here is the latest research on allergy and asthma.

ASBMB Publications

The American Society for Biochemistry and Molecular Biology (ASBMB) includes the Journal of Biological Chemistry, Molecular & Cellular Proteomics, and the Journal of Lipid Research. Discover the latest research from ASBMB here.