Verfügbarkeit: Intranuclear quality control compartment (INQ)

Intranuclear quality control compartment (INQ): mechanism of misfolded protein sorting and physiological function

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Bibliographische Detailangaben
1. Verfasser:	Khokhrina, Maria (VerfasserIn)
Format:	Abschlussarbeit Buch
Sprache:	English
Veröffentlicht:	Heidelberg [2018]
Schlagworte:	Hochschulschrift
Online-Zugang:	Inhaltsverzeichnis Inhaltsverzeichnis
Beschreibung:	Enthält 2 Beiträge aus verschiedenen Zeitschriften
Beschreibung:	vi, 142 Seiten, 36 verschieden gezählte Seiten Illustrationen, Diagramme 30 cm

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Datensatz im Suchindex

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adam_text	TABLE OF CONTENTS 1. INTRODUCTION.............................................................................................................................. 1 1.1 P ROTEIN Q UALITY C ONTROL NETWORK IN THE CYTOSOL ............................................. 1.1.1 PROTEIN FOLDING AND MISFOLDING O F NEWLY SYNTHETIZED PEPTIDES AT THE RIBOSOME 1.1.2 RIBOSOME QUALITY CONTROL PATHWAY (RQC) ......................................................... 1.1.3 MOLECULAR CHAPERONES ........................................................................................ 1.1.4 DEGRADATION O F MISFOLDEDPROTEINS AND AGGREGATE CLEARANCE ................................ 13 1.1.5 SPATIAL PROTEIN QUALITY CONTROL .................................................................................. 15 1.2 I NTERPLAY BETWEEN CYTOSOLIC AND NUCLEAR QUALITY CONTROL ................................20 1.2.1 INQ (INTRANUCLEAR QUALITY CONTROL COMPARTMENT) .................................................... 20 1.3 N UCLEUS AS A SITE OF MULTIFUNCTIONAL QUALITY CONTROL ........................................... 22 1.3.1 DNA DAMAGE SIGNALING ORCHESTRATED BY NUCLEAR BODIES ........................................ 23 2. A IM S ....................................................................................................................................................25 3. R E SU L T S...........................................................................................................................................26 3.1. P RINCIPLES OF MISFOLDED PROTEIN TARGETING TO INQ .................................................... 26 3.1.1 UBIQUITINATION IS NOT AN ESSENTIAL SIGNAL FO R INQ SORTING ..................................... 26 3.1.2 EFFECT O F STIL ON THE NUCLEAR SORTING O F MISFOLDED PROTEINS .................................... 29 3.1.3 EFFECT O FS IS L ON THE NUCLEAR SORTING O F MISFOLDED PROTEINS ................................... 31 3.2 A PPROACHES TO IDENTIFY NOVEL INQ TARGETING FACTORS AND INQ SUBSTRATES ..... 34 3.2.1 IMMUNOPRECIPITATION WITH BTN2 BINDER ..................................................................... 36 3.2.2 FLUORESCENT SORTING O F NUCLEAR AGGREGATES ............................................................... 40 3.2.3 AGGREGATE PELLETING .................................................................................................... 43 3.2.4 IDENTIFICATION O F PROTEINS INTERACTING WITH BTN2 UPON HEAT STRESS VIA IN VIVO CROSS- LINKING AND AFFINITY PURIFICATION ......................................................................................... 45 3.3 INQ AS A GENOTOXIC STRESS RESPONSE COMPARTMENT .................................................... 51 3.3.1 GENOTOXIC STRESS TRIGGERS INQ FORMATION .................................................................. 51 3.3.2 INQ PLAYS A ROLE IN THE S PHASE CHECKPOINT CONTROL. ............................................... 58 3.3.3 INQ IS INVOLVED IN DNA REPLICATION CHECKPOINT ...................................................... 62 3.3.4 M RCL RE-LOCALIZES AT INQ UPON GENOTOXIC STRESS ..................................................... 63 3.3.5 DNA CHECKPOINT RECOVERY IS IMPEDED IN BTN2A DIA2A CELLS ..................................... 65 3.3.6 TARGETING HSP42 TO THE NUCLEUS DOES NOT FUNCTIONALLY REPLACE BTN2 UPON GENOTOXIC STRESS IN BTN2A DIA2A CELLS .................................................................................................. 67 3.4 R OLE OF AGGREGASES B TN 2, H SP 42 AND H SP 40 S ISL IN RIBOSOME QUALITY CONTROL PATHWAY ..............................................................................................................................................73 4. D ISC U SSIO N ....................................................................................................................................76 4.1 S ORTING SIGNALS AND FACTORS OF MISFOLDED PROTEIN SEQUESTRATION ...................... 76 4.2 E XPERIMENTAL APPROACHES TO IDENTIFY INQ SUBSTRATES AND ADDITIONAL SORTING FACTORS ...............................................................................................................................................79 4.3 P HYSIOLOGICAL FUNCTION OF B TN 2 ( AND INQ) UPON HEAT SHOCK ..................................82 4.4 R OLE OF B TN 2- DRIVEN INQ IN CHECKPOINT CONTROL DURING GENOTOXIC STRESS ...... 83 4.5 CAT- TAILS : AN INHERENT AGGREGATION SIGNAL THAT ALLOWS PROTEIN SEQUESTRATION INDEPENDENT FROM AGGREGASE FUNCTION ....................................................86 5. M ATERIALS AND M E T H O D S.............................................................................................. 88 5.1 S OFTWARE AND EQUIPMENT ...................................................................................................... 88 5.2 M ICROSCOPE ................................................................................................................................. 90 5.3 E XPENDABLE ITEM S .....................................................................................................................91 -FCX UO UJ CONTENTS 5.4 C HEMICALS .....................................................................................................................................92 5.5 Y EAST METHODS ..........................................................................................................................100 5.5.1 TRANSFORMATION O F YEA ST ........................................................................................... 100 5.5.2 GENOMIC MANIPULATION O F YE A S T .............................................................................. 100 5.5.3 EXTRACTION O F GENOMIC DNA FROM YEA ST .................................................................. 101 5.5.5 GROWTH CONDITIONS .................................................................................................... 102 5.5.6 IN VIVO CROSSLINKING ................................................................ 103 5.5 .7 AGGREGATE PELLETING ASSAY ........................................................................................ 103 5.5.8 CO-IMMUNIPRECIPITATION ........................................................................................... 105 5.5.9 FLOW CYTOMETRY ANALYSIS O F CELLULAR AGGREGATES .................................................... 106 5.5.10 YEAST CELL CYCLE PROGRESSION ANALYSIS .................................................................... 107 5.5.11 YEAST DNA CONTENT ANALYSIS ................................................................................... 107 5.6 M OLECULAR B IOLOGY M ETHODS ............................................................................................ 108 5.6.1 POLYMERASE CHASE REACTION (PCR) ......................................................................... 108 5.6.2 AGAROSE GEL ELECTROPHORESIS ................................................................................... 108 5.6.3 RESTRICTION DIGEST O F DN A ........................................................................................ 109 5.6.4 TRANSFORMATION OFE . COLI ......................................................................................... 109 5.7 B IOCHEMICAL M ETH O D S ........................................................................................................... 110 5.7.1 DETERMINATION O F PROTEIN CONCENTRATION ................................................................. 110 5.7.2 PROTEIN PRECIPITATION WITH TRICHLOROACETIC ACID (TCA) .......................................... 110 5.7.3 SDSPOLYACRYLAMIDE GEL ELECTROPHORESIS (SDS-PAGE) ......................................... 110 5.7.4 WESTERN BLOT, IMMUNOSTAINING AND DETECTION O F PROTEIN BANDS ............................. I L L 5.7.5 SILVER STAINING ........................................................................................................... 112 5.7.6 COUPLING O F BTN2 ANTIBODIES TO SEPHAROSE BEADS .................................................... 113 5.7.7 PURIFICATION O F BTN2-HBH AFTER CROSSLINKING .......................................................... 113 5.8 M ICROSCOPY ............................................................................................................................... 116 5.8.1 FIXATION AND DNA STAINING FO R THE WIDE FIE LD MICROSCOPY .................................... 116 5.8.2 IMAGE ACQUISITION, PROCESSING AND DATA ANALYSIS ................................................... 116 5.8.3 IMMUNOFLUORESCENCE MICROSCOPY ............................................................................. 116 5.9 O THER METH O DS .........................................................................................................................117 5.9.1 PROTEIN IDENTIFICATION BY MASS-SPECTROMETRY .......................................................... 117 LIST OF FIGURES.................................................................................................................118 LIST OF TABLES................................................................................................................... 119 BIBLIOGRAPHY.................................................................................................................... 120 ACKNOWLEDGEMENTS................................................................................................... 141
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author	Khokhrina, Maria
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spelling	Khokhrina, Maria Verfasser (DE-588)1153085976 aut Intranuclear quality control compartment (INQ) mechanism of misfolded protein sorting and physiological function presented by Maria Khokhrina, Master of Biology Heidelberg [2018] vi, 142 Seiten, 36 verschieden gezählte Seiten Illustrationen, Diagramme 30 cm txt rdacontent n rdamedia nc rdacarrier Enthält 2 Beiträge aus verschiedenen Zeitschriften Dissertation Ruperto-Carola University of Heidelberg 2018 Mit einer Zusammenfassung in englischer und deutscher Sprache (DE-588)4113937-9 Hochschulschrift gnd-content B:DE-101 application/pdf http://d-nb.info/1154329550/04 Inhaltsverzeichnis DNB Datenaustausch application/pdf http://bvbr.bib-bvb.de:8991/F?func=service&doc_library=BVB01&local_base=BVB01&doc_number=030665665&sequence=000001&line_number=0001&func_code=DB_RECORDS&service_type=MEDIA Inhaltsverzeichnis
spellingShingle	Khokhrina, Maria Intranuclear quality control compartment (INQ) mechanism of misfolded protein sorting and physiological function
subject_GND	(DE-588)4113937-9
title	Intranuclear quality control compartment (INQ) mechanism of misfolded protein sorting and physiological function
title_auth	Intranuclear quality control compartment (INQ) mechanism of misfolded protein sorting and physiological function
title_exact_search	Intranuclear quality control compartment (INQ) mechanism of misfolded protein sorting and physiological function
title_full	Intranuclear quality control compartment (INQ) mechanism of misfolded protein sorting and physiological function presented by Maria Khokhrina, Master of Biology
title_fullStr	Intranuclear quality control compartment (INQ) mechanism of misfolded protein sorting and physiological function presented by Maria Khokhrina, Master of Biology
title_full_unstemmed	Intranuclear quality control compartment (INQ) mechanism of misfolded protein sorting and physiological function presented by Maria Khokhrina, Master of Biology
title_short	Intranuclear quality control compartment (INQ)
title_sort	intranuclear quality control compartment inq mechanism of misfolded protein sorting and physiological function
title_sub	mechanism of misfolded protein sorting and physiological function
topic_facet	Hochschulschrift
url	http://d-nb.info/1154329550/04 http://bvbr.bib-bvb.de:8991/F?func=service&doc_library=BVB01&local_base=BVB01&doc_number=030665665&sequence=000001&line_number=0001&func_code=DB_RECORDS&service_type=MEDIA
work_keys_str_mv	AT khokhrinamaria intranuclearqualitycontrolcompartmentinqmechanismofmisfoldedproteinsortingandphysiologicalfunction

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