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
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.
dc.description.sponsorship Brandeis University, Graduate School of Arts and Sciences
dc.format.mimetype application/pdf
dc.language English
dc.language.iso eng
dc.publisher Brandeis University
dc.relation.ispartofseries Brandeis University Theses and Dissertations
dc.rights Copyright by Zhi Yang 2015
dc.subject spliceosome
dc.subject U1 snRNP
dc.subject gold nanoparticle
dc.subject electron microscopy
dc.subject EM reconstruction
dc.subject S. cerevisiae U1 snRNP
dc.title Development of A Site-specific Affinity Pin to Label Subunits in the S. cerevisiae U1 snRNP for Structural Studies
dc.type Thesis
dc.contributor.department Department of Biochemistry
dc.degree.name MS
dc.degree.level Masters
dc.degree.discipline Biochemistry
dc.degree.grantor Brandeis University, Graduate School of Arts and Sciences


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