The RNA-binding protein YT521-B plays a role in the alternative splice site selection and is regulated by tyrosine phosphorylation:
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| Main Author: | |
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| Format: | Thesis Book |
| Language: | English German |
| Published: |
2005
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| Subjects: | |
| Online Access: | Inhaltsverzeichnis |
| Item Description: | Erlangen-Nürnberg, Univ, Diss., 2005. - Enth. dt. Zsfassung |
| Physical Description: | 135 S. Ill., graph. Darst. |
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_______ Contents CONTENTS
CONTENTS __|
ABBREVIATIONS Iv
ZUSAMMENFASSUNG vii
ABSTRACT viii
1. INTRODUCTION __ 1
1.1. Pre mRNA splicing /
1.1.1. Basic splicing reaction and spliceosome assembly 1
1.1.2. Exon recognition and intron bridging 4
1.1.3. Alternative splicing 6
1.1.4. Regulation of the alternative splicing 8
1.1.4.1. Phosphorylation dependent control of the pre mRNA splicing machinery 9
1.1.5. Function of alternative splicing 10
1.1.5.1. Introduction of stop codons 11
1.1.5.2. Changes of the protein structure 12
1.1.5.3. Changes of the mRNA properties 14
1.2. RNA interacting domains 14
1.2.1. RNA recognition motif (RRM) 16
1.2.2. K homology (KH) domain 16
1.2.3. Double stranded RNA binding domain (dsRBD) 17
1.3. Nuclear organization and subnuclear bodies 18
1.4. YT521 B as a putative splicing factor 20
2. RESEARCH OVERVIEW 11
3. MATERIALS AND METHODS 23
3.1. Materials 23
3.1.1. Chemicals 23
3.1.2. Enzymes 24
3.1.3. Cell lines and media 24
3.1.4. Bacterial strains and media 24
3.1.5. Antibiotics 24
3.1.6. Antibodies 25
3.1.7. Plasmids 25
3.1.7.1. Clones from the lab collection or outside sources 25
3.1.7.2. Newly made clones 26
3.1.8. Primers 26
3.1.8.1. Primers used for huYT52! B gene 26
3.1.8.2. Primers used for cloning 27
3.1.8.3. Primers used for mutagenesis 27
3.1.8.4. Primers used for RT PCR 28
3.2. Methods__ 28
3.2.1. PCR amplification of DNA_ 28
3.2.2. Plasmid DNA isolation 29
3.2.3. Determination of nucleic acid concentration 29
3
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3.3A tin iWrarwrt ¦wnpn.ril of UNA ; »
3JL7. Preparation of punt g.td» iriU . — 31
3i«. Tmufcnnatioa of £cott caDa 31
3J.9.Ez|acMiaDMd|MrificMioaariflS«tsripniMiHJa£co« 3?
3iI0. Detcrmmat* of pratefe ooaocatntioa ___— 33
3.ZH.hhicfck*ck) online aaaay . 33
3.2.12. RNA electrophontic nobility dull tatty (RNA get ahift) 33
3.2.l2.!.IUdioitbeliD|ofRNAoligaaacleotidet ___ M
32.13. Freein».th.wm»md«*aitomg of euk«yoticc «» M
JJ.U.Trmribakmof.nAMyoticcdl. 35
3J.15.Fixingin»dKdeuk»yo«icceU.oiicowrflipf ., 35
3il6. bnmuooOaining 36
3Xll. Quantification of ccO ittiii«ig__ 36
3.Z18.tannunopreeipil«kaofpralni*_. 36
3.2.19. Solubility «s»»y 37
3.2^0 lsohlkin of micleaprotemotlrattind RNA imanMMMecHiiuaion 38
3^.21. Ekctroohoremof BrotriM 39
3.2.22. Stammg of protein gel* 39
3.2.23. Western Wotting 40
3.2.24. In Who fpKchg ascty 40
3.2.25. Isolation of total RNA 41
3.2.26. RT PCR 42
3.3. Databases and computational tools 42
4. RESULTS 44
4.1. Human YT521 B gene structure ; 44
4.2. YT52I B protein has a modular structure 49
4.3. YTH: a new domain in mir tvar prnteivts 34
4.4. Antiserum specific for YTH domain ** ,
4.5. TH ANLS4 YT521 B deletion mutant binds topurine rich RNA 60
4.6. Mutations within YTH domain changes localization ofYT521 B 64
4.7. Mutation within YTH domain change ability of YT521 B to influence CD44 alternative pre mRNA
i • 66
splicing ^_ —— ——— 4.8. YT521 B is phosphorylatedon tyrosine residues . 67
4.9. The proline rich region is required for YT521 B phosphorylation 7/
4.10. YT521 B binds to c Abl kinase 12
4.11. YT521 B colocalizes with c Abl in the nucleus 73
4.12. YT521 B shuttles between nucleus and cytosol 7^
4.13. YT bodies disperse after tyrosine phosphorylation 7S
4.14. The solubility ofYT521 B is regulated by phosphorylation ____ 77
4. IS. YT521 B regulates the splicing of several minigenes in a concentration dependent manner 78
4.16. YT521 B acts on the interleukin 4 receptor pre mRNA —. *
4.17. Phosphorylation changes the ability of YT52I B to influence splice site selection oflL 4R pre
mRNA —— — *3
ii
Contents 4.18. Phosphorylalion changes the ability ofYTS21 B to influence CD44 alternative pre mRNA splicing,
but does not affect the splicing pattern oftra2 beta and E1A pre mRNAs 87
5. DISCUSSION 89
5.1. YT521 B orthologs and YTH containing proteins 89
5.2. YTH is a possible novel nucleic acid binding domain 91
5.3. Tyrosine phosphorylation regulates the nuclear localization ofYT521 B 93
5.4. Similarities between effects qftyrosine phosphorylation in the cell membrane and the nucleus 95
5.5. Splice site selection is regulated by tyrosine phosphorylation 95
5.6. Working model of YT521 B function 97
REFERENCES 99
iii
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| dewey-full | 572.865 |
| dewey-hundreds | 500 - Natural sciences and mathematics |
| dewey-ones | 572 - Biochemistry |
| dewey-raw | 572.865 |
| dewey-search | 572.865 |
| dewey-sort | 3572.865 |
| dewey-tens | 570 - Biology |
| discipline | Biologie |
| format | Thesis Book |
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| indexdate | 2024-07-09T20:08:01Z |
| institution | BVB |
| language | English German |
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| spelling | Rafalska, Ilona 1976- Verfasser (DE-588)130157384 aut The RNA-binding protein YT521-B plays a role in the alternative splice site selection and is regulated by tyrosine phosphorylation von Ilona Rafalska 2005 135 S. Ill., graph. Darst. txt rdacontent n rdamedia nc rdacarrier Erlangen-Nürnberg, Univ, Diss., 2005. - Enth. dt. Zsfassung Erlangen-Nürnberg, Univ., Diss., 2005 Tyrosin (DE-588)4186526-1 gnd rswk-swf RNS-Spleißen (DE-588)4201877-8 gnd rswk-swf Genregulation (DE-588)4122166-7 gnd rswk-swf Genexpression (DE-588)4020136-3 gnd rswk-swf Kernproteine (DE-588)4389511-6 gnd rswk-swf Phosphorylierung (DE-588)4174459-7 gnd rswk-swf (DE-588)4113937-9 Hochschulschrift gnd-content Genexpression (DE-588)4020136-3 s Genregulation (DE-588)4122166-7 s RNS-Spleißen (DE-588)4201877-8 s Kernproteine (DE-588)4389511-6 s Tyrosin (DE-588)4186526-1 s Phosphorylierung (DE-588)4174459-7 s b DE-604 HBZ Datenaustausch application/pdf http://bvbr.bib-bvb.de:8991/F?func=service&doc_library=BVB01&local_base=BVB01&doc_number=013194124&sequence=000002&line_number=0001&func_code=DB_RECORDS&service_type=MEDIA Inhaltsverzeichnis |
| spellingShingle | Rafalska, Ilona 1976- The RNA-binding protein YT521-B plays a role in the alternative splice site selection and is regulated by tyrosine phosphorylation Tyrosin (DE-588)4186526-1 gnd RNS-Spleißen (DE-588)4201877-8 gnd Genregulation (DE-588)4122166-7 gnd Genexpression (DE-588)4020136-3 gnd Kernproteine (DE-588)4389511-6 gnd Phosphorylierung (DE-588)4174459-7 gnd |
| subject_GND | (DE-588)4186526-1 (DE-588)4201877-8 (DE-588)4122166-7 (DE-588)4020136-3 (DE-588)4389511-6 (DE-588)4174459-7 (DE-588)4113937-9 |
| title | The RNA-binding protein YT521-B plays a role in the alternative splice site selection and is regulated by tyrosine phosphorylation |
| title_auth | The RNA-binding protein YT521-B plays a role in the alternative splice site selection and is regulated by tyrosine phosphorylation |
| title_exact_search | The RNA-binding protein YT521-B plays a role in the alternative splice site selection and is regulated by tyrosine phosphorylation |
| title_full | The RNA-binding protein YT521-B plays a role in the alternative splice site selection and is regulated by tyrosine phosphorylation von Ilona Rafalska |
| title_fullStr | The RNA-binding protein YT521-B plays a role in the alternative splice site selection and is regulated by tyrosine phosphorylation von Ilona Rafalska |
| title_full_unstemmed | The RNA-binding protein YT521-B plays a role in the alternative splice site selection and is regulated by tyrosine phosphorylation von Ilona Rafalska |
| title_short | The RNA-binding protein YT521-B plays a role in the alternative splice site selection and is regulated by tyrosine phosphorylation |
| title_sort | the rna binding protein yt521 b plays a role in the alternative splice site selection and is regulated by tyrosine phosphorylation |
| topic | Tyrosin (DE-588)4186526-1 gnd RNS-Spleißen (DE-588)4201877-8 gnd Genregulation (DE-588)4122166-7 gnd Genexpression (DE-588)4020136-3 gnd Kernproteine (DE-588)4389511-6 gnd Phosphorylierung (DE-588)4174459-7 gnd |
| topic_facet | Tyrosin RNS-Spleißen Genregulation Genexpression Kernproteine Phosphorylierung Hochschulschrift |
| url | http://bvbr.bib-bvb.de:8991/F?func=service&doc_library=BVB01&local_base=BVB01&doc_number=013194124&sequence=000002&line_number=0001&func_code=DB_RECORDS&service_type=MEDIA |
| work_keys_str_mv | AT rafalskailona thernabindingproteinyt521bplaysaroleinthealternativesplicesiteselectionandisregulatedbytyrosinephosphorylation |