RGD Reference Report - Substrate specificity plays an important role in uncoupling the catalytic and scaffolding activities of rat testis DNA topoisomerase IIalpha. - Rat Genome Database

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Substrate specificity plays an important role in uncoupling the catalytic and scaffolding activities of rat testis DNA topoisomerase IIalpha.

Authors: Bakshi, R  Galande, S  Muniyappa, K 
Citation: Bakshi R, etal., J Biomol Struct Dyn. 2001 Apr;18(5):749-60.
RGD ID: 1580614
Pubmed: PMID:11334111   (View Abstract at PubMed)
DOI: DOI:10.1080/07391102.2001.10506704   (Journal Full-text)

Topoisomerase II (topo II) is a dyadic enzyme found in all eukaryotic cells. Topo II is involved in a number of cellular processes related to DNA metabolism, including DNA replication, recombination and the maintenance of genomic stability. We discovered a correlation between the development of postnatal testis and increased binding of topo IIalpha to the chromatin fraction. We used this observation to characterize DNA-binding specificity and catalytic properties of purified testis topo IIalpha. The results indicate that topo IIalpha binds a substrate containing the preferred site with greater affinity and, consequently, catalyzes the conversion of form I to form IV DNA more efficiently in contrast to substrates lacking such a site. Interestingly, topo IIalpha displayed high-affinity and cooperativity in binding to the scaffold associated region. In contrast to the preferred site, however, high-affinity binding of topo IIalpha to the scaffold-associated region failed to result in enhanced catalytic activity. Intriguingly, competition assays involving scaffold-associated region revealed an additional DNA-binding site within the dyadic topo IIalpha. These results implicate a dual role for topo IIalpha in vivo consistent with the notion that its sequestration to the chromatin might play a role in chromosome condensation and decondensation during spermatogenesis.



Gene Ontology Annotations    Click to see Annotation Detail View

Biological Process

  
Object SymbolSpeciesTermQualifierEvidenceWithNotesSourceOriginal Reference(s)
Top2aRatDNA topological change  IDA  RGD 

Cellular Component

  
Object SymbolSpeciesTermQualifierEvidenceWithNotesSourceOriginal Reference(s)
Top2aRatDNA topoisomerase type II (double strand cut, ATP-hydrolyzing) complex  IDA  RGD 

Molecular Function

  
Object SymbolSpeciesTermQualifierEvidenceWithNotesSourceOriginal Reference(s)
Top2aRatATP binding  IDA  RGD 
Top2aRatchromatin binding  IDA  RGD 
Top2aRatDNA binding  IDA  RGD 
Top2aRatDNA topoisomerase activity  IDA  RGD 
Top2aRatDNA topoisomerase type II (double strand cut, ATP-hydrolyzing) activity  IDA  RGD 
Top2aRatsequence-specific DNA binding  IDA  RGD 

Objects Annotated

Genes (Rattus norvegicus)
Top2a  (DNA topoisomerase II alpha)


Additional Information