Kinetic study of the DNA annealing properties of RECQ5β helicase

RecQ family helicases are critical for maintaining genomic integrity. Many RecQ family helicases not only unwind duplex, and other more complicated DNA structures, but also possess, interestingly, DNA annealing （strand pairing） activity. Here, we sys- tematically investigated the DNA annealing prope...

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Bibliographic Details
Published in:Chinese science bulletin 2012-04, Vol.57 (11), p.1280-1287
Main Authors: Ding, XiuYan, Xu, YaNan, Li, Wei, Wang, PengYe, Xi, XuGuang, Dou, ShuoXing
Format: Article
Language:English
Subjects:
adenosine diphosphate
Chemistry/Food Science
dissociation
DNA annealing
DNA结合
DNA结构
Earth Sciences
Engineering
Humanities and Social Sciences
Life Sciences
multidisciplinary
Physics
Science
Science (multidisciplinary)
single-stranded DNA
催化机制
动力学性质
单链DNA
属性
解旋酶
退火
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Summary:RecQ family helicases are critical for maintaining genomic integrity. Many RecQ family helicases not only unwind duplex, and other more complicated DNA structures, but also possess, interestingly, DNA annealing （strand pairing） activity. Here, we sys- tematically investigated the DNA annealing properties of RECQ513 by measuring DNA annealing kinetics, equilibrium DNA binding, and kinetics of dissociation from ssDNA. RECQ513 catalyzed DNA annealing most efficiently when the enzyme mole- cules covered approximately 40%-50% of the DNA strand, in the absence or presence of different nucleotide cofactors （AMPPNP ATPγS, or ADP） under our buffer conditions. A comparative study with RECQ5β^1-662 confirmed that the C-terminal region of RECQ5[3 was essential for its high DNA annealing activity. These results contribute to our understanding of the mechanism of DNA annealing catalyzed by RecQ family helicases.
ISSN:1001-6538
1861-9541
DOI:10.1007/s11434-012-4981-x