Genetic analyses of kindlins in mice [Elektronische Ressource] / Siegfried Ussar

Siegfried Ussar

Sujets

• Biologie
• Gesundheit

Informations

• Publié par: ludwig-maximilians-universitat_munchen
• Publié le: 01 janvier 2008
• Nombre de lectures: 17
• Langue: Deutsch
• Poids de l'ouvrage: 14 Mo

Extrait

Dissertation zur Erlangung des Doktorgrades
der Fakultät für Chemie und Pharmazie der
Ludwig-Maximilians-Universität München






Genetic analyses of Kindlins in mice



Siegfried Ussar

aus

Graz, Österreich

2008
Erklärung
Diese Dissertation wurde im Sinne von §13 Abs. 3 der Promotionsordnung vom 29. Januar
1998 von Herrn Prof. Dr. Reinhard Fässler betreut.



Ehrenwörtliche Versicherung

Diese Dissertation wurde selbstständig, ohne unerlaubte Hilfe erarbeitet.



München, am 19.09.2008







(Siegfried Usar)







Dissertation eingereicht am: 22.09.08
1. Gutachter Prof. Dr. Reinhard Fässler
2. Gutachter PD Dr. Klaus-Peter Janssen

Mündliche Prüfung am: 23.10.2008 Table of Contents
Table of Contents
TABLE OF CONTENTS............................................................................................................I
AIM OF THE THESIS...............................................................................................................1
LIST OF PUBLICATIONS........................................................................................................2
SUMMARY...............................................................................................................................3
INTRODUCTION......................................................................................................................6
KINDLINS.6
Kindlin gene and protein structure ...................................................................................................... 7
FERM domains 8
PH/PTB superfamily........................................................................................................................ 8
The role of UNC-112 in C. elegans....................................................................................................... 9
Integrins................................................................................................................................................ 11
Integrin function in vivo...................................................................................................................... 13
Murine pre- and peri-implantation development ............................................................................... 13
The role of integrins during pre- and peri-implantation development........................................... 14
Skin.................... 15
The role of integrins in the epidermis............................................................................................ 16
Intestine.............. 17
The role of integrins in the intestine.............................................................................................. 18
Platelets............................................................................................................................................. 19
The role of integrins in platelets.................................................................................................... 21
Integrin activation (Inside-out signalling)........................................................................................... 22
Talin mediates integrin activation ................................................................................................. 23
Outside-in Signalling ......................................................................................................................... 25
Kindlin function in mammalian cells 26
Kindler Syndrome.............................................................................................................................. 26
Kindlins in integrin signalling............................................................................................................. 28
PAPER SUMMARIES ............................................................................................................31
ITable of Contents
The Kindlins: subcellular localization and expression during murine development................... 31
Kindlin-2 controls bidirectional signalling of integrins................................................................... 32
Integrin-linked kinase: integrin's mysterious partner ..................................................................... 33
Kindlin-3 is essential for integrin activation and platelet aggregation.......................................... 34
SILAC-mouse for quantitative proteomics uncovers Kindlin-3 as an essential factor for red
blood cell function............................................................................................................................... 35
Leukocyte Adhesion Deficiency-III (LAD-III) is caused by mutations in the adhesion protein
Kindlin-3, * equal contribution ........................................................................................................... 36
Loss of Kindlin-1 causes skin atrophy and early postnatal ulcerative colitis .............................. 37
Colocalization of kindlin-1, kindlin-2 and migfilin at keratinocyte focal contacts and relevance
to the pathophysiology of Kindler syndrome................................................................................... 38
Expression of truncated kindlin-1 leads to abnormal adhesion and migration of keratinocytes39
ACKNOWLEDGEMENTS......................................................................................................40
CURRICULUM VITAE ...........................................................................................................41
REFERENCES.......................................................................................................................43
APPENDIX.............................................................................................................................51
IIAim of the thesis
Aim of the thesis
The function of the three Kindlin proteins is unknown. Recently, mutations in the human
FERMT1 gene, coding for the Kindlin-1 protein, were shown to cause Kindler Syndrome, a
rare autosomal recessive skin blistering disease. In addition to the skin blister phenotype,
several other abnormalities have been reported, although a clear genotype to phenotype
correlation is not evident. Cell-culture based work on Kindlin-1 and Kindlin-2 demonstrated
an important role of these proteins in integrin-mediated cell adhesion and adhesion
strengthening by the actin cytoskeleton. No functional data on Kindlin-3 have been reported,
although the high homology to the other Kindlins suggests that they share similar functions,
at least at integrin adhesion sites.
The overall goal of this thesis was to describe the consequences of loss of each Kindlin
protein in mice. Four major aims were defined for addressing these tasks; first,
characterisation of the spatial and temporal expression pattern of the three Kindlin mRNAs
and proteins. Second, generation of mouse strains deficient for each Kindlin. Third, analyses
of their phenotypes and comparison of the Kindlin-1 knockout phenotype in mice with the
human Kindler Syndrome. Fourth, identification of the molecular basis for the phenotypes.

1List of Publications
List of Publications
This thesis is based on the following publications, which are referred to in the text by their
Roman numerals (I-IX):

I Ussar S, Wang HV, Linder S, Fässler R, Moser M (2006) The Kindlins: subcellular
localization and expression during murine development. Exp Cell Res 312: 3142-
3151.

II Montanez E*, Ussar S*, Schifferer M, Bösl M, Zent R, Moser M, Fässler R (2008)
Kindlin-2 controls bidirectional signaling of integrins. Genes Dev 22: 1325-1330. *
equal contribution

III Grashoff C, Thievessen I, Lorenz K, Ussar S, and Fässler R (2004) Integrin-linked
kinase: integrin's mysterious partner. Current opinion in cell biology 16(5), 565-571

IV Moser M, Nieswandt B, Ussar S, Pozgajova M, Fässler R (2008) Kindlin-3 is
essential for integrin activation and platelet aggregation. Nat Med 14: 325-330.

V Kruger M*, Moser M*, Ussar S, Thievessen I, Luber C, A, Forner F, Schmidt S,
Zanivan S, Fässler R, Mann M, (2008) SILAC mouse for quantitative proteomics
uncovers kindlin-3 as an essential factor for red blood cell function. Cell 134: 353-
364. * equal contribution

VI. Svensson L*, Howarth K*, McDowall A, Patzak I, Evans R, Ussar S, Metin A, Fried
M, Tomlinson I,Hogg N, (submitted) Leukocyte Adhesion Deficiency-III (LAD-III) is
caused by mutations in the adhesion protein Kindlin-3, * equal contribution

VII Ussar S, Moser M, Widmaier M, Rognoni E, Harrer C, Genzel-Boroviczeny O,
Fässler R (submitted) Loss of Kindlin-1 causes skin atrophy and lethal neonatal
ulcerative colitis.

VIII Lai-Cheong JE, Ussar S, Arita K, Hart IR, McGrath JA (2008) Colocalization of
kindlin-1, kindlin-2, and migfilin at keratinocyte focal adhesion and relevance to the
pathophysiology of Kindler syndrome. J Invest Dermatol 128: 2156-2165.

IX Has C, Ludwig RJ, Herz C, Kern JS, Ussar S, Ochsendorf FR, Kaufmann