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Purification of UV Cross-Linked RNA-Protein Complexes by Phenol-Toluol Extraction.

Record Type:	Language materials, printed : Monograph/item
Title/Author:	Purification of UV Cross-Linked RNA-Protein Complexes by Phenol-Toluol Extraction./
Author:	Urdaneta Zurbaran, Erika Cristina.
Published:	Ann Arbor : ProQuest Dissertations & Theses, : 2020,
Description:	141 p.
Notes:	Source: Dissertations Abstracts International, Volume: 81-11, Section: B.
Contained By:	Dissertations Abstracts International81-11B.
Subject:	Biology. -
Online resource:	http://pqdd.sinica.edu.tw/twdaoapp/servlet/advanced?query=27991984
ISBN:	9798643189329

Purification of UV Cross-Linked RNA-Protein Complexes by Phenol-Toluol Extraction.
Urdaneta Zurbaran, Erika Cristina.

Purification of UV Cross-Linked RNA-Protein Complexes by Phenol-Toluol Extraction. - Ann Arbor : ProQuest Dissertations & Theses, 2020 - 141 p.

Source: Dissertations Abstracts International, Volume: 81-11, Section: B.

Thesis (Ph.D.)--Humboldt Universitaet zu Berlin (Germany), 2020.

This item must not be sold to any third party vendors.

RNA binding proteins play key functions in post-transcriptional regulation of gene expression. By binding to RNA, they control RNA editing, transport, stability and translation. In the last decade, significant advances have been made in the elucidation of bacterial post-transcriptional mechanisms. It is becoming increasingly clear that this layer of regulation also plays an important role in pathogenesis and antibiotic resistance. The analysis of RNA-protein complexes (RNPs) at the proteome level has been driven by the (m)RNA interactome capture technology which isolates the proteome cross-linked to poly-adenylated (m)RNA. This has resulted in the identification of hundreds of novel RBPs in a diversity of eukaryotic species ranging from humans to yeast. However, poly-adenylation is absent in functional RNA from bacteria and other classes of -eukaryotic- regulatory RNAs. This work was aimed to overcome that limitation by developing a novel and unbiased method for the purification of UV-cross-linked RNPs in living cells: PTex (Phenol Toluol extraction). The purification principle is solely based on physicochemical properties of cross-linked RNPs versus unbound proteins or RNA, and it is impartial towards specific RNA or proteins; enabling for the first time a system-wide analysis of non-poly(A) RNA interacting proteins in both eukaryotic (HEK293) and prokaryotic (Salmonella Typhimurium) cells.

ISBN: 9798643189329Subjects--Topical Terms:

599573
Biology.
Subjects--Index Terms:

RNA-protein complexes

Purification of UV Cross-Linked RNA-Protein Complexes by Phenol-Toluol Extraction.
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245 1 0 $a Purification of UV Cross-Linked RNA-Protein Complexes by Phenol-Toluol Extraction.
260 1 $a Ann Arbor : $b ProQuest Dissertations & Theses, $c 2020
300 $a 141 p.
500 $a Source: Dissertations Abstracts International, Volume: 81-11, Section: B.
500 $a Advisor: Selbach , Matthias;Ringrose , Leonie;Beckmann , Benedikt.
502 $a Thesis (Ph.D.)--Humboldt Universitaet zu Berlin (Germany), 2020.
506 $a This item must not be sold to any third party vendors.
520 $a RNA binding proteins play key functions in post-transcriptional regulation of gene expression. By binding to RNA, they control RNA editing, transport, stability and translation. In the last decade, significant advances have been made in the elucidation of bacterial post-transcriptional mechanisms. It is becoming increasingly clear that this layer of regulation also plays an important role in pathogenesis and antibiotic resistance. The analysis of RNA-protein complexes (RNPs) at the proteome level has been driven by the (m)RNA interactome capture technology which isolates the proteome cross-linked to poly-adenylated (m)RNA. This has resulted in the identification of hundreds of novel RBPs in a diversity of eukaryotic species ranging from humans to yeast. However, poly-adenylation is absent in functional RNA from bacteria and other classes of -eukaryotic- regulatory RNAs. This work was aimed to overcome that limitation by developing a novel and unbiased method for the purification of UV-cross-linked RNPs in living cells: PTex (Phenol Toluol extraction). The purification principle is solely based on physicochemical properties of cross-linked RNPs versus unbound proteins or RNA, and it is impartial towards specific RNA or proteins; enabling for the first time a system-wide analysis of non-poly(A) RNA interacting proteins in both eukaryotic (HEK293) and prokaryotic (Salmonella Typhimurium) cells.
520 $a RNA-Bindungsproteine spielen Schlusselfunktionen bei der post-transkriptionellen Regulation der Genexpression. Durch Bindung an RNA steuern sie die RNA-Aufbereitung, den Transport, die Stabilitat und die Translation. In den letzten zehn Jahren wurden bedeutende Fortschritte bei der Aufklarung bakterieller post-transkriptioneller Mechanismen erzielt. Es wird immer deutlicher, dass diese Regulierungsebene auch bei der Pathogenese und Antibiotikaresistenz eine wichtige Rolle spielt. Die Analyse von RNA-Protein-Komplexen (RNPs) auf Proteomebene wurde durch die (m)RNA-interactome-capture Technologie vorangetrieben, die den Teil des Proteoms isoliert, welcher mit polyadenylierter (m)RNA vernetzt ist. Dies hat zur Identifizierung von Hunderten von neuen RBPs in einer Vielzahl von eukaryontischen Arten, vom Menschen bis zur Hefe, gefuhrt. Allerdings fehlt die Poly-Adenylierung in der funktionellen RNA von Bakterien und anderen Klassen von -eukaryontischen- regulatorischen RNAs. Ziel dieser Arbeit war es, diese Einschrankung durch die Entwicklung einer neuartigen und unvoreingenommenen Methode zur Aufreinigung von UV-vernetzten RNPs in lebenden Zellen zu uberwinden: PTex (Phenol-Toluol-Extraktion). Das Reinigungsprinzip basiert ausschlieslich auf den physikalisch-chemischen Eigenschaften von vernetzten RNPs gegenuber ungebundenen Proteinen oder RNA; es ist dabei unparteiisch gegenuber spezifischen RNAs oder Proteinen und ermoglicht somit erstmals eine systemweite Analyse von nicht-poly-(A)-RNA-interagierenden Proteinen sowohl in eukaryontischen (HEK293) als auch in prokaryontischen (Salmonella Typhimurium) Zellen.
590 $a School code: 5416.
650 4 $a Biology. $3 599573
653 $a RNA-protein complexes
653 $a Phenol-toluol extraction
690 $a 0306
710 2 $a Humboldt Universitaet zu Berlin (Germany). $3 1335299
773 0 $t Dissertations Abstracts International $g 81-11B.
790 $a 5416
791 $a Ph.D.
792 $a 2020
793 $a English
856 4 0 $u http://pqdd.sinica.edu.tw/twdaoapp/servlet/advanced?query=27991984