Development of A Site-specific Affinity Pin to Label Subunits in the S. cerevisiae U1 snRNP for Structural Studies

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dc.contributor.advisor Krummel, Daniel Pomeranz en_US
dc.contributor.author Yang, Zhi
dc.date.accessioned 2015-05-18T16:26:17Z
dc.date.available 2015-05-18T16:26:17Z
dc.date.issued 2015
dc.identifier.uri http://hdl.handle.net/10192/30585
dc.description.abstract Eukaryotic protein coding gene precursor-mRNA transcripts are processed or spliced to remove the non-coding sequences (introns) and connect or splice together the coding sequences (exons) for translation. The splicing process is catalyzed by a complex and dynamic macromolecular assembly called the spliceosome. Integral to the spliceosome are five uridine-rich small nuclear ribonucleoprotein particles, the U1, U2, U4, U5, and U6 snRNPs. The three-dimensional structures for most of the U snRNPs remain unclear. Recently, a low-resolution (~25 angstrom) electron microscopy negative-stain reconstruction of the S. cerevisiae 18-subunit U1 snRNP (~800kDa) as well as an improved resolution (~16 angstroms) electron microscopy reconstruction of the particle in vitreous ice (cryo) were obtained. However, at such resolutions interpretation of the experimental density map and thus inference into the function of individual subunits is a formidable challenge. Herein, I present strategies designed to aid model building into these low- resolution electron microscopy reconstructions. Specifically, I report on progress made to both label the single RNA subunit of the budding yeast U1 snRNP (U1 snRNA) and a protein subunit with a monodisperse gold nanoparticle. en_US
dc.description.sponsorship Brandeis University, Graduate School of Arts and Sciences en_US
dc.format.mimetype application/pdf en_US
dc.language English en_US
dc.language.iso eng en_US
dc.publisher Brandeis University en_US
dc.rights Copyright by Zhi Yang 2015 en_US
dc.subject spliceosome en_US
dc.subject U1 snRNP en_US
dc.subject gold nanoparticle en_US
dc.subject electron microscopy en_US
dc.subject EM reconstruction en_US
dc.subject S. cerevisiae U1 snRNP en_US
dc.title Development of A Site-specific Affinity Pin to Label Subunits in the S. cerevisiae U1 snRNP for Structural Studies en_US
dc.type Thesis en_US
dc.contributor.department Department of Biochemistry en_US
dc.degree.name MS en_US
dc.degree.level Masters en_US
dc.degree.discipline Biochemistry en_US
dc.degree.grantor Brandeis University, Graduate School of Arts and Sciences en_US


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