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dc.contributor.advisor Hsieh, Tao-shih en_US
dc.contributor.author Capp, Christopher Lee en_US
dc.date.accessioned 2011-05-20T19:33:48Z
dc.date.available 2011-05-20T19:33:48Z
dc.date.issued 2011 en_US
dc.identifier.uri http://hdl.handle.net/10161/3814
dc.description Dissertation en_US
dc.description.abstract <p>RecQ4, a member of the conserved RecQ family of helicases, is involved in replication and associated with several clinical syndromes. Although biologically important, the biochemistry of RecQ4 has remained elusive. We have expressed and purified Drosophila melanogaster RecQ4 from a baculovirus expression system. Biochemical characterization of the helicase, ATP hydrolysis, annealing, and binding activities of the enzyme has been performed, using native and non-native gel electrophoresis and thin layer chromatography, among other techniques. These reveal that RecQ4 is a 3' to 5' helicase that is stimulated by the presence of single-stranded DNA 3' of the duplex DNA region to be unwound. The enzyme is also capable of annealing complementary DNA strands, though this is inhibited by AMPPNP, a non-hydrolyzable analog of ATP. RecQ4 also forms a stable complex with single-stranded DNA in the presence of AMPPNP. We argue that the helicase activity of RecQ4 is important to the process of DNA replication. This leads to the conclusion that two helicases, RecQ4 and the Mcm2-7 complex, are involved in replication. The manner of their simultaneous involvement is not intuitive, and so models by which the two enzymes may cooperate are discussed.</p> en_US
dc.subject Biochemistry en_US
dc.subject DNA replication en_US
dc.subject Drosophila en_US
dc.subject helicase en_US
dc.subject RecQ en_US
dc.title The Biochemical Characterization of Drosophila melanogaster RecQ4 Helicase en_US
dc.type Dissertation en_US
dc.department Biochemistry en_US

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