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Spliceosomal small nuclear ribonucleoprotein particle trafficking and maturation in the nuclear compartment

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Title: Spliceosomal small nuclear ribonucleoprotein particle trafficking and maturation in the nuclear compartment
Author(s): Patel, Snehal B.
Director of Research: Bellini, Michel
Doctoral Committee Chair(s): Bellini, Michel
Doctoral Committee Member(s): Gennis, Robert B.; Kannanganattu, Prasanth; Nair, Satish K.; Shapiro, David J.
Department / Program: Biochemistry
Discipline: Biochemistry
Degree Granting Institution: University of Illinois at Urbana-Champaign
Degree: Ph.D.
Genre: Dissertation
Subject(s): splicing small nuclear ribonucleoproteins (snRNP) oocyte post-transcriptional gene regulation
Abstract: We show for the first time that upon injection into the cytoplasm of the oocyte, fluorescein-labeled spliceosomal snRNAs, in the context of functional snRNPs, are targeted to elongating pre-mRNAs. This finding presents us with a novel assay with which to dissect the mechanism by which snRNPs are targeted to nascent pre-mRNA transcripts. Two critical advantages offered by this system are immediately evident. First, it allows us to investigate the mechanisms employed to recruit snRNPs as it actually transpires within the realm of the cell nucleus. Second, it allows a genome-wide analysis of snRNP recruitment to nascent transcripts, and, hence, the conclusions drawn from these studies do not depend on the sequence of any particular promoter or pre-mRNA. Indeed, it is with this assay that we have stumbled upon a most unanticipated discovery: Contrary to the current paradigm, the co-transcriptional recruitment of splicing snRNPs to nascent transcripts is not contingent on their role in splicing in vivo. Based on these and other data, we have constructed a two-step recruitment-loading model wherein snRNPs are first recruited to pre-mRNA transcripts and only then loaded directly onto cis-acting sequences on nascent pre-mRNA. While conducting studies on snRNP trafficking, a new discovery was made. We found that the lampbrush chromosomes could be visualized by light microscopy in vivo, and that these chromosomes have an architecture that is identical with those in formaldehyde treated nuclear spread preparations. Importantly, we now have the first system with which we can examine the dynamic interactions of macromolecules with specific RNA polymerase II transcriptional units in the live nucleus.
Issue Date: 2011-01-14
Rights Information: Copyright 2009 Snehal Patel
Date Available in IDEALS: 2011-01-14
Date Deposited: December 2

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