Domains of CstF-64 and their functions in polyadenylation

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dc.contributor.committeeChairMacDonald, Clinton C.
dc.contributor.committeeMemberWhelly, Sandra M.
dc.contributor.committeeMemberSridhara, S.
dc.contributor.committeeMemberThomas, Jeffrey
dc.contributor.committeeMemberFaust, Charles
dc.creatorHockert, John Andrew
dc.date.accessioned2016-11-14T23:21:12Z
dc.date.available2011-02-18T21:21:00Z
dc.date.available2016-11-14T23:21:12Z
dc.date.issued2007-12
dc.degree.departmentTTUHSC -- Cell Biology and Biochemistryen_US
dc.description.abstractPolyadenylation, a critical process for expression of most eukaryotic genes, requires multiple protein factors and pre-mRNA elements. However, the essential nature of polyadenylation proteins precludes in vivo determination of their precise functions. We present a straightforward, sensitive, and adaptable in vivo polyadenylation assay, the stem-loop luciferase assay for polyadenylation (SLAP) as a tool to dissect the functions of critical polyadenylation proteins. Our investigation focused on the CstF-64 subunit of the cleavage stimulation factor (CstF), which binds to the pre-mRNA downstream of the site of cleavage. Using an mRNA with a modified downstream element requiring co-expression of an MS2-CstF-64 fusion protein, we determined that the RNA binding domain (RBD), Hinge, and C-terminal domains (CTD) of CstF-64 were indispensable for polyadenylation in vivo. Furthermore, we showed that the Hinge domain was required for CstF-64 nuclear localization and CstF-77 association, suggesting that CstF complex formation and nuclear import are essential steps in polyadenylation.
dc.format.mimetypeapplication/pdf
dc.identifier.urihttp://hdl.handle.net/2346/15998en_US
dc.language.isoeng
dc.publisherTexas Tech Universityen_US
dc.rights.availabilityUnrestricted.
dc.subjectPolyadenylationen_US
dc.subjectCleavage stimulation factor (CstF)en_US
dc.titleDomains of CstF-64 and their functions in polyadenylation
dc.typeDissertation

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