The snoRNA HBII-52 absent in Prader-Willi syndrome patients regulates the alternative splicing of the serotonin receptor 5-HT_1tn2_1tnCR [Elektronische Ressource] / vorgelegt von Shivendra Kishore

Shivendra2

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• Gesundheit

Informations

• Publié par: friedrich-alexander-universitat_erlangen-nurnberg
• Publié le: 01 janvier 2007
• Nombre de lectures: 7
• Langue: English
• Poids de l'ouvrage: 2 Mo

Extrait

The snoRNA HBII-52 absent in Prader-Willi Syndrome
patients regulates the alternative splicing of the serotonin
receptor 5-HT R 2C





Den Naturwissenschaftlichen Fakultäten
der Friedrich-Alexander-Universität Erlangen-Nürnberg
zur
Erlangung des Doktorgrades





vorgelegt von
Shivendra Kishore
aus Patna, Indien



2006


Als Dissertation genehmigt von den Naturwissenschaftlichen Fakultäten
Der Universität Erlangen-Nürnberg.













Tag der mündlichen Prüfung: 27 März 2006


Vorsitzender der Prüfungskomission: Prof. Dr. Donat-Peter Häder
Erstberichterstatter: PD Dr. Fritz Titgemeyer
Zweitberichterstatter: Prof. Dr. Michael Wegner








To see a World in a Grain of Sand
And a Heaven in a Wild Flower
Hold Infinity in the palm of your hand
And Eternity in an hour.



William Blake
‘Auguries of Innocence’,1794









ACKNOWLEDGMENTS
This work, like most others reflects in its utmost simplicity the positives coming out of a long
arduous effort. What it fails to reveal, are the feelings attached to the periods of continued mental
conflicts, indecisiveness and despair. Also, it conceals within, the spurts of hopes and excitements at every
small accomplishement, the comfort of being together in times of desperation and the bliss of unraveling
new horizons. There are many people to whom I owe the fulfillment of this dissertation.

My sincerest gratitude goes to Prof. Stefan Stamm for his confidence in me all these years and in
helping me work freely on topics that were immensely challenging. I shall never cease to admire his
contribution in bringing out the inquisitiveness within me. His insight into things, presented before me a
vast gradient of knowledge that always kept me on my toes. I owe him most for his patience and
understanding. Without him, I find it difficult to imagine myself having grown to such levels of maturity
and understanding.

I would also like to thank the former and present members of the Stamms lab, Peter Stoilov,
Marieta Gencheva, Ilona Rafalska, Bettina Heinrich, Tatyana Novoyatleva, Zhaiyi Zhang, Natalya
Benderska and Dominique Olbert for their enthusiasm and constructive criticisms. My special thanks go to
Yesheng Tang for his rational advice, moral support and for providing congenial atmosphere for open
discussions. His contribution in making me think and work out strategies will always remain
unparalleled.

I am also thankful to all the members of the Institute of Biochemistry, in particular, Prof. Wegner
and his group for freely sharing their resources with us.

Many thanks are due to Prof. Alexander Hüttenhofer and Patrice Vitali for their critical remarks
and insightful suggestions about snoRNAs during the initial stages. I am thankful to Prof. Ingmar
Blümke and Prof. Winfried Neuhuber for their generous help in brain dissections.

I appreciate my friends Shobhit, Stefanie, Gagan, Kunal and Aditya for their memorable company
in Germany and unconditional support in times when I needed most. Thanks to Anurag and Kapil for
staying with me through my ups and downs for last 8 years. I am grateful to my sister for instilling in me
courage to strive for higher goals in life.

In the end, I would like to thank my parents and all my teachers who have believed in me and
have been the milestones of my life. Besides failures in which I remember them, I am sure; there will be
successes in which they will walk with me.






PUBLICATIONS
Parts of this work are included in the following publications:

Kishore S and Stamm S. (2006) The snoRNA HBII-52 regulates alternative splicing of the
serotonin receptor 2C, Science, Jan 13;311(5758):230-2.



Other publications as a part of the doctoral work:

Bergin A.M., Balder B., Kishore S., Swärd K., Hahn-Zoric M., Löwenhagen O., Hanson
L.A. and Padyukov L. Common variations in the IL4R gene affect splicing and influence
natural expression of the soluble isoform. (in submission)

Tang Y., Novoyatleva T., Benderska N., Kishore S., Thanaraj A. and Stamm S. (2005)
Analysis of Alternative Splicing In Vivo using Minigenes. Handbook of RNA Biochemistry,
WILEY-VCH Verlag GmbH & Co. KHaA, Weinheim.






Contents
CONTENTS
ABBREVIATIONS................................................................................................................... iv
ZUSAMMENFASSUNG........................................................................................................... vi
ABSTRACT............................................................................................................................. vii
1. INTRODUCTION.................................................................................................................. 1
1.1. Pre-mRNA Splicing.....................................................................................................................2
1.1.1. Exon recognition and intron bridging..................................................................................5
1.1.2. Alternative splicing..............................................................................................................7
1.1.3. Regulation of alternative splicing........................................................................................8
1.1.4. Function of alternative splicing ...........................................................................................9
1.2. RNA editing...............................................................................................................................13
1.3. Regulatory role of non-coding RNAs15
1.3.1. Small nucleolar RNAs (or snoRNAs)................................................................................16
1.3.2. Structure and function of methylation guide C/D box snoRNA........................................17
1.3.3. Structure and function of pseudouridylation guide H/ACA box snoRNA ........................19
1.3.4. Biogenesis and assembly of snoRNP particles ..................................................................20
1.3.5. Associated proteins............................................................................................................21
1.3.6. Cellular RNA targets for eukaryotic snoRNA guides........................................................22
1.3.7. Brain specific snoRNAs and genomic imprinting .............................................................23
1.4. Serotonin 5-HT receptors..........................................................................................................25
1.4.1. Human Serotonin receptor 2C (5-HT R) .........................................................................26 2C
1.4.2. 5-HT R: Genomic structure and its alternative spliced variants ......................................27 2C
1.4.3 R editing by ADARs and its functional implication .............................................. 28 2C
1.5. Prader-Willi syndrome (PWS) ..................................................................................................31
1.5.1. Inheritance in PWS ............................................................................................................31
1.5.2. Imprinted PWS domain .....................................................................................................32
1.5.3. snoRNAs and PWS............................................................................................................34
2. RESEARCH OVERVIEW................................................................................................... 36
3. MATERIALS AND METHODS .......................................................................................... 37
3.1. Materials...................................................................................................................................37
3.1.1. Chemicals ..........................................................................................................................37
3.1.2. Enzymes............37
3.1.3. Cell lines and media...........................................................................................................38
3.1.4. Bacterial strains and media................................................................................................38
3.1.5. Antibiotics..........................................................................................................................39
3.1.6. Antibodies..........................................................................................................................39
3.1.7. Plasmids.............................................................................................................................39
3.1.8. Primers..............42
3.1.9. Brain Tissues .......................................................................................