Holdings: The role of translational velocity in mutant protein folding

The role of translational velocity in mutant protein folding: how CFTR translation modulates its folding

Saved in:

Bibliographic Details
Main Author:	Rauscher, Robert (Author)
Format:	Thesis Book
Language:	English
Published:	Hamburg [2018]
Subjects:	Hochschulschrift
Online Access:	Inhaltsverzeichnis Inhaltsverzeichnis
Physical Description:	96, XVII Seiten Illustrationen, Diagramme 21 cm

Staff View

MARC


LEADER	00000nam a2200000 c 4500
001	BV045440363
003	DE-604
005	20190828
007	t
008	190204s2018 gw a\|\|\| m\|\|\| 00\|\|\| eng d
015			\|a 18,H09 \|2 dnb
016	7		\|a 1163625795 \|2 DE-101
035			\|a (OCoLC)1118983588
035			\|a (DE-599)DNB1163625795
040			\|a DE-604 \|b ger \|e rda
041	0		\|a eng
044			\|a gw \|c XA-DE
049			\|a DE-83 \|a DE-355
084			\|a 570 \|2 sdnb
100	1		\|a Rauscher, Robert \|e Verfasser \|0 (DE-588)1147688109 \|4 aut
245	1	0	\|a The role of translational velocity in mutant protein folding \|b how CFTR translation modulates its folding \|c vorgelegt von Robert Rauscher, M.Sc.
264		1	\|a Hamburg \|c [2018]
300			\|a 96, XVII Seiten \|b Illustrationen, Diagramme \|c 21 cm
336			\|b txt \|2 rdacontent
337			\|b n \|2 rdamedia
338			\|b nc \|2 rdacarrier
502			\|b Dissertation \|c Universität Hamburg \|d 2018
655		7	\|0 (DE-588)4113937-9 \|a Hochschulschrift \|2 gnd-content
856	4	2	\|m B:DE-101 \|q application/pdf \|u http://d-nb.info/1163625795/04 \|3 Inhaltsverzeichnis
856	4	2	\|m DNB Datenaustausch \|q application/pdf \|u http://bvbr.bib-bvb.de:8991/F?func=service&doc_library=BVB01&local_base=BVB01&doc_number=030825887&sequence=000001&line_number=0001&func_code=DB_RECORDS&service_type=MEDIA \|3 Inhaltsverzeichnis
999			\|a oai:aleph.bib-bvb.de:BVB01-030825887

Record in the Search Index

_version_	1804179334996426752
adam_text	TABLE OF CONTENTS PUBLICATIONS................................................................................................. II TABLE OF CONTENTS....................................................................................ILL LIST OF FIGURES........................................................................................... VI LIST OF TABLES........................................................................................... VIII LIST OF ABBREVIATIONS..............................................................................IX ZUSAMMENFASSUNG ........................................................ XI ABSTRACT......................................................................................................XII 1. INTRODUCTION..........................................................................................1 1.1. TRANSLATION........................................................................................................................ 1 1.1.1. THE PHASES OF EUKARYOTIC TRANSLATION....................................................................... 1 1.1.2. FEATURES THAT MODULATE TRANSLATIONAL EFFICIENCY AND LOCAL VELOCITY ........................ 3 1.1.3. COTRANSLATIONAL PROTEIN FOLDING.................................................................................. 6 1.2. THE CYSTIC FIBROSIS TRANSMEMBRANE CONDUCTANCE REGULATOR ............................... 8 1.2.1. CFTR BIOGENESIS AND FOLDING ................................................................................... 8 1.2.2. CFTR FUNCTION..........................................................................................................10 1.2.3. MUTATIONS IN C FTR ................................................................................................... 11 1.2.4. CYSTIC FIBROSIS AND CFTR RELATED DISORDERS ........................................................ 13 1.2.5. DEGRADATION OF MISFOLDED CFTR .............................................................................. 14 1.2.6. PHARMACOLOGICAL TREATMENT OF CYSTIC FIBROSIS ....................................................... 16 1.3. ER-STRESS AND THE UNFOLDED PROTEIN RESPONSE .................................. 17 1.4. AIM OF THIS THESIS...........................................................................................................20 2. RESULTS....................................... . ...................................................... - 21 2.1. SEQUENCE FEATURES INFLUENCE TRANSLATIONAL VELOCITY.................................................21 2.1.1. CFTR TRANSLATION VELOCITY IS NOT UNIFORM ................................................................21 2.1.2. MEASURES OF TRANSLATIONAL VELOCITY BARELY CORRELATE WITH ONE ANOTHER .................. 22 2.2. TOWARDS DEEPER COVERAGE OF SPECIFIC TRANSCRIPTS IN RIBOSEQ EXPERIMENTS ...... 25 2.3. TRANSLATIONAL VELOCITY IS EMBEDDED INTO THE CFTR TRANSCRIPT .............................. 28 2.4. EFFECTS OF ALTERED TRANSLATIONAL VELOCITY ALONG THE CFTR TRANSCRIPT .................... 30 2.4.1. SSNPS OCCUR AT DIFFERENT FREQUENCIES.....................................................................30 2.4.2. MUTATIONS DIFFERENTIALLY AFFECT CFTR STEADY-STATE EXPRESSION AND MATURATION... 32 2.4.3. THE SSNP T2562G DIFFERENTIALLY AFFECTS WILD-TYPE AND MUTANT CFTR VARIANTS .33 2.4.4. THE T2562G SSNP AMELIORATES CFTR ACTIVITY OF SPECIFIC CFTR VARIANTS ......... 36 2.4.5. THE T2562G SSNP AUGMENTS STABILITY AND MUTANT CFTR VARIANTS ..................... 38 2.4.6. THE T2562G SSNP ENHANCED THE EFFECT OF CORRECTOR ON CFTR VARIANTS WITH CF- CAUSING M UTATION.......................................................................................................................40 2.5. MECHANISTIC INSIGHTS INTO RPL12 KNOCK-DOWN MEDIATED CFTR RESCUE .............. 42 2.6. CFTR MISFOLDING INDUCES TRANSCRIPTOME CHANGES..................................................46 3. DISCUSSION..............................................................................................52 3.1. TRANSLATIONAL DYNAMICS IN CFTR FOLDING....................................................................52 3.1.1. CFTR BIOSYNTHESIS IS NON-UNIFORM.....................................................................52 3.1.2. IMPERFECT CORRELATION OF VARIOUS MEASURES OF TRANSLATION VELOCITY ............... 53 3.1.3. TERIBO-SEQ PROVIDES INFORMATION ON RIBOSOME TRAFFIC FOR INDIVIDUAL TRANSCRIPTS................................................................................................................................ 54 3.1.4. TRANSLATIONAL SPEED IS EMBEDDED IN THE CFTR TRANSCRIPT...............................55 3.2. SSNPS FREQUENCIES IN PATIENTS DEVIATE FROM THESE IN HEALTHY POPULATION .......... 56 3.3. THE T2562G SSNP DIFFERENTIALLY AFFECTS DIFFERENT CFTR VARIANTS WITH A MAJOR CF MUTATION................................................................................................................ 57 3.4. RPL12-MEDIATED CODON-SPECIFIC TRANSLATIONAL SLOW-DOWN RESHAPES THE CFTR TRANSLATIONAL LANDSCAPE.............................................................................................................. 59 3.5. CFTR MISFOLDING COMPROMISES THE STRESS RESPONSE ............................................. 60 4. MATERIAL AND METHODS .................................................................... 63 4.1. MATERIALS............................................................................................................................63 4.1.1. REAGENTS AND CHEMICALS.........................................................................................63 4.1.2. ENZYMES AND BUFFERS............................................................................................... 63 4.1.3. ANTIBODIES ................................................. 64 4.1.4. KITS............................................................................................................................. 64 4.1.5. OLIGONUCLEOTIDES ................................................................... 65 4.1.6. BUFFERS AND REAGENTS 69 4.2. METHODS.................................................... 72 4.2.1. CELL CULTURE............................................................................................................... 72 4.2.2. RIBOSOME PROFILING AND MRNA SEQUENCING .......................................................... 72 4.3. BIOCHEMICAL EXPERIMENTS ............................................................................................. 75 4.3.1. CLONING AND TRANSFECTION......................................................................................... 75 4.3.2. WESTERN BLOTTING.......................................................................................................76 4.3.3. ISOLATION OF GENOMIC DNA......................................... 76 4.3.4. SPLICING ASSAY.......................................................................................................... 77 4.3.5. THERMAL AGGREGATION PROPENSITY ASSAY..................................................................77 4.3.6. PARTIAL PERMEABILIZATION OF THE CELLS AND LIMITED PROTEOLYSIS ................................. 77 4.3.7. ACTIVITY MEASUREMENTS............................................................................................ 78 4.3.6. SHORT-CIRCUIT CURRENT (LSC) MEASUREMENT...................................................................78 4.3.9. TOTAL RNA EXTRACTION, CDNA SYNTHESIS AND QUANTITATIVE PC R .............................. 78 4.3.10. TRNA TAILORED MICROARRAYS........................................................................................79 LIST OF REFERENCES ............... 80 SUPPLEMENTARY INFORMATION...............................................................92 LIST OF HAZARDOUS SUBSTANCES USED IN THE STUDY .................. XII ACKNOWLEDGEMENTS..............................................................................XVI DECLARATION ON OATH .......................................................................... XVII
any_adam_object	1
author	Rauscher, Robert
author_GND	(DE-588)1147688109
author_facet	Rauscher, Robert
author_role	aut
author_sort	Rauscher, Robert
author_variant	r r rr
building	Verbundindex
bvnumber	BV045440363
ctrlnum	(OCoLC)1118983588 (DE-599)DNB1163625795
discipline	Biologie
format	Thesis Book
fullrecord	<?xml version="1.0" encoding="UTF-8"?><collection xmlns="http://www.loc.gov/MARC21/slim"><record><leader>01370nam a2200337 c 4500</leader><controlfield tag="001">BV045440363</controlfield><controlfield tag="003">DE-604</controlfield><controlfield tag="005">20190828 </controlfield><controlfield tag="007">t</controlfield><controlfield tag="008">190204s2018 gw a\|\|\| m\|\|\| 00\|\|\| eng d</controlfield><datafield tag="015" ind1=" " ind2=" "><subfield code="a">18,H09</subfield><subfield code="2">dnb</subfield></datafield><datafield tag="016" ind1="7" ind2=" "><subfield code="a">1163625795</subfield><subfield code="2">DE-101</subfield></datafield><datafield tag="035" ind1=" " ind2=" "><subfield code="a">(OCoLC)1118983588</subfield></datafield><datafield tag="035" ind1=" " ind2=" "><subfield code="a">(DE-599)DNB1163625795</subfield></datafield><datafield tag="040" ind1=" " ind2=" "><subfield code="a">DE-604</subfield><subfield code="b">ger</subfield><subfield code="e">rda</subfield></datafield><datafield tag="041" ind1="0" ind2=" "><subfield code="a">eng</subfield></datafield><datafield tag="044" ind1=" " ind2=" "><subfield code="a">gw</subfield><subfield code="c">XA-DE</subfield></datafield><datafield tag="049" ind1=" " ind2=" "><subfield code="a">DE-83</subfield><subfield code="a">DE-355</subfield></datafield><datafield tag="084" ind1=" " ind2=" "><subfield code="a">570</subfield><subfield code="2">sdnb</subfield></datafield><datafield tag="100" ind1="1" ind2=" "><subfield code="a">Rauscher, Robert</subfield><subfield code="e">Verfasser</subfield><subfield code="0">(DE-588)1147688109</subfield><subfield code="4">aut</subfield></datafield><datafield tag="245" ind1="1" ind2="0"><subfield code="a">The role of translational velocity in mutant protein folding</subfield><subfield code="b">how CFTR translation modulates its folding</subfield><subfield code="c">vorgelegt von Robert Rauscher, M.Sc.</subfield></datafield><datafield tag="264" ind1=" " ind2="1"><subfield code="a">Hamburg</subfield><subfield code="c">[2018]</subfield></datafield><datafield tag="300" ind1=" " ind2=" "><subfield code="a">96, XVII Seiten</subfield><subfield code="b">Illustrationen, Diagramme</subfield><subfield code="c">21 cm</subfield></datafield><datafield tag="336" ind1=" " ind2=" "><subfield code="b">txt</subfield><subfield code="2">rdacontent</subfield></datafield><datafield tag="337" ind1=" " ind2=" "><subfield code="b">n</subfield><subfield code="2">rdamedia</subfield></datafield><datafield tag="338" ind1=" " ind2=" "><subfield code="b">nc</subfield><subfield code="2">rdacarrier</subfield></datafield><datafield tag="502" ind1=" " ind2=" "><subfield code="b">Dissertation</subfield><subfield code="c">Universität Hamburg</subfield><subfield code="d">2018</subfield></datafield><datafield tag="655" ind1=" " ind2="7"><subfield code="0">(DE-588)4113937-9</subfield><subfield code="a">Hochschulschrift</subfield><subfield code="2">gnd-content</subfield></datafield><datafield tag="856" ind1="4" ind2="2"><subfield code="m">B:DE-101</subfield><subfield code="q">application/pdf</subfield><subfield code="u">http://d-nb.info/1163625795/04</subfield><subfield code="3">Inhaltsverzeichnis</subfield></datafield><datafield tag="856" ind1="4" ind2="2"><subfield code="m">DNB Datenaustausch</subfield><subfield code="q">application/pdf</subfield><subfield code="u">http://bvbr.bib-bvb.de:8991/F?func=service&doc_library=BVB01&local_base=BVB01&doc_number=030825887&sequence=000001&line_number=0001&func_code=DB_RECORDS&service_type=MEDIA</subfield><subfield code="3">Inhaltsverzeichnis</subfield></datafield><datafield tag="999" ind1=" " ind2=" "><subfield code="a">oai:aleph.bib-bvb.de:BVB01-030825887</subfield></datafield></record></collection>
genre	(DE-588)4113937-9 Hochschulschrift gnd-content
genre_facet	Hochschulschrift
id	DE-604.BV045440363
illustrated	Illustrated
indexdate	2024-07-10T08:18:13Z
institution	BVB
language	English
oai_aleph_id	oai:aleph.bib-bvb.de:BVB01-030825887
oclc_num	1118983588
open_access_boolean
owner	DE-83 DE-355 DE-BY-UBR
owner_facet	DE-83 DE-355 DE-BY-UBR
physical	96, XVII Seiten Illustrationen, Diagramme 21 cm
publishDate	2018
publishDateSearch	2018
publishDateSort	2018
record_format	marc
spelling	Rauscher, Robert Verfasser (DE-588)1147688109 aut The role of translational velocity in mutant protein folding how CFTR translation modulates its folding vorgelegt von Robert Rauscher, M.Sc. Hamburg [2018] 96, XVII Seiten Illustrationen, Diagramme 21 cm txt rdacontent n rdamedia nc rdacarrier Dissertation Universität Hamburg 2018 (DE-588)4113937-9 Hochschulschrift gnd-content B:DE-101 application/pdf http://d-nb.info/1163625795/04 Inhaltsverzeichnis DNB Datenaustausch application/pdf http://bvbr.bib-bvb.de:8991/F?func=service&doc_library=BVB01&local_base=BVB01&doc_number=030825887&sequence=000001&line_number=0001&func_code=DB_RECORDS&service_type=MEDIA Inhaltsverzeichnis
spellingShingle	Rauscher, Robert The role of translational velocity in mutant protein folding how CFTR translation modulates its folding
subject_GND	(DE-588)4113937-9
title	The role of translational velocity in mutant protein folding how CFTR translation modulates its folding
title_auth	The role of translational velocity in mutant protein folding how CFTR translation modulates its folding
title_exact_search	The role of translational velocity in mutant protein folding how CFTR translation modulates its folding
title_full	The role of translational velocity in mutant protein folding how CFTR translation modulates its folding vorgelegt von Robert Rauscher, M.Sc.
title_fullStr	The role of translational velocity in mutant protein folding how CFTR translation modulates its folding vorgelegt von Robert Rauscher, M.Sc.
title_full_unstemmed	The role of translational velocity in mutant protein folding how CFTR translation modulates its folding vorgelegt von Robert Rauscher, M.Sc.
title_short	The role of translational velocity in mutant protein folding
title_sort	the role of translational velocity in mutant protein folding how cftr translation modulates its folding
title_sub	how CFTR translation modulates its folding
topic_facet	Hochschulschrift
url	http://d-nb.info/1163625795/04 http://bvbr.bib-bvb.de:8991/F?func=service&doc_library=BVB01&local_base=BVB01&doc_number=030825887&sequence=000001&line_number=0001&func_code=DB_RECORDS&service_type=MEDIA
work_keys_str_mv	AT rauscherrobert theroleoftranslationalvelocityinmutantproteinfoldinghowcftrtranslationmodulatesitsfolding

Holdings

There is no print copy available.

Interlibrary loan Place Request Caution: Not in THWS collection! Indexes
Indexes

MARC

Record in the Search Index

There is no print copy available.

Similar Items