The fission yeast Aurora kinase: possible mechanisms of activation and its role in regulating microtubule dynamics
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| Format: | Thesis Book |
| Language: | English |
| Published: |
2010
|
| Subjects: | |
| Online Access: | Inhaltsverzeichnis |
| Physical Description: | IX, 111 S. Ill., graph. Darst. |
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| 100 | 1 | |a Huang, Yu-Hua |d 1975- |e Verfasser |0 (DE-588)142810592 |4 aut | |
| 245 | 1 | 0 | |a The fission yeast Aurora kinase |b possible mechanisms of activation and its role in regulating microtubule dynamics |c von Yu-Hua Huang |
| 264 | 1 | |c 2010 | |
| 300 | |a IX, 111 S. |b Ill., graph. Darst. | ||
| 336 | |b txt |2 rdacontent | ||
| 337 | |b n |2 rdamedia | ||
| 338 | |b nc |2 rdacarrier | ||
| 502 | |a Tübingen, Univ., Diss., 2010 | ||
| 655 | 7 | |0 (DE-588)4113937-9 |a Hochschulschrift |2 gnd-content | |
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| 999 | |a oai:aleph.bib-bvb.de:BVB01-020880166 | ||
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| adam_text | TABLE
OF
CONTENTS
ACKNOWLEDGEMENTS
.............................................................................................................
I
LIST
OF
FIGURES
........................................................................................................................
V
ABBREVIATIONS
......................................................................................................................VII
SUMMARY
AND
OUTLOOK
...................................................................................................
IX
1
GENERAL
INTRODUCTION
................................................................................................
1
1.1
C
ELL
CYCLE
.........................................................................................................................
2
1.2
M
ITOSIS
................................................................................................................................
4
1.2.1
THE
MITOTIC
SPINDLE
.......................................................................................................
5
1.2.2
KINETOCHORE-MICROTUBULE
ATTACHMENT
.........................................................................
7
1.2.3
MICROTUBULE-ASSOCIATED
PROTEINS
(MAPS)
AND
MOTOR
PROTEINS
ACTING
DURING
MITOSIS
9
1.3
M
ITOTIC
FUNCTIONS
OF
THE
A
URORA
B
KINASE
...........................................................
12
1.3.1
CHROMOSOME
CONDENSATION
AND
HETEROCHROMATIN
MODIFICATION
............................
12
1.3.2
CORRECTION
OF
ABERRANT
ATTACHMENT
...........................................................................
14
1.3.3
SPINDLE
ASSEMBLY
CHECKPOINT
(SA
C)
........................................................................
18
1.3.4
CYTOKINESIS
AND
ABSCISSION
........................................................................................
19
1.4
THE
REGULATION
OF
AURORA
B
.....................................................................................
20
1.5
FISSION
YEAST
AS
A
MODEL
ORGANISM
..........................................................................
22
1.6
AIM
OF
THIS
THESIS
..........................................................................................................
23
2
THE
REGULATION
OF
THE
ARK1
COMPLEX
IN
MITOSIS
.................................
24
2.1
INTRODUCTION
..................................................................................................................
24
2.2
R
ESULTS
............................................................................................................................
25
2.2.1
CAFFEINE
SENSITIVITY
IS
INFLUENCED
BY
MUTATION
OF
ADE6
............................................
25
2.2.2
STRAINS
EXPRESSING
ARKL-T7
AND-T11
ARE
CAFFEINE
SENSITIVE
..................................
26
2.2.3
CAFFEINE
REDUCES
HISTONE
H3
SERINE
10
PHOSPHORYLATION
(H3S10PH)
....................
28
2.2.4
DELETION
OF
CHKL
OR
RAD3
DOES
NOT
AFFECT
HISTONE
H3
SERINE
10
PHOSPHORYLATION
(H3S10PH)
...............................................................................................................................
31
2.2.5
ARKL
KINASE
ACTIVITY
IS
DECREASED
IN
THE
PRESENCE
OF
CAFFEINE
................................32
2.3
DISCUSSION
......................................................................................................................
34
2.3.1
ADENINE
AND
CAFFEINE
..................................................................................................34
2.3.2
HOW
DOES
CAFFEINE
INFLUENCE
ARKL
ACTIVITY?
.............................................................35
2.4
S
UPPLEMENTAL
DATA
.....................................................................................................
39
II
3
PHOSPHORYLATION
BY
THE
AURORA
KINASE
ARK1
INHIBITS
THE
DEPOLYMERIZATION-INDUCING
ACTIVITY
OF
KINESIN-8
_________________
41
3.1
INTRODUCTION
..................................................................................................................
41
3.2
C
ONTRIBUTIONS
................................................................................................................
44
3.3
R
ESULTS
.............................................................................................................................
45
3.3
.
/
ARKL
PHOSPHORYLATES
SEVERAL
KINETOCHORE
PROTEINS
IN
VITRO
..............................
45
33.2
ARKL
PHOSPHORYLATES
KINESIN-8
(KLP5
AND
KLP6)
IN
VITRO
......................................
47
333
ARKL
PHOSPHORYLATES
SEVERAL
RESIDUES
WITHIN
KLP5/KLP6
IN
VITRO
.........................
49
33.4
KLP
5
IS
PHOSPHORYLATED
ON
SEVERAL
SITES
DURING
MITOSIS
IN
VIVO
.............................
52
3.3.5
CELLS
EXPRESSING
KLP5-T224D
ARE
SIMILARLY
RESISTANT
TO
A
MT-DESTABILIZING
DRUG
AS
KLP5K
CELLS
............................................................................................................................
53
3.3.6
CELLS
EXPRESSING
KLP5-T224D
CELLS
SHOW
A
RETENTION
OF
THE
SAC
PROTEIN
BUBL,
BUT
NOT
MAD2
AT
KINETOCHORES
DURING
PROMETAPHASE,
SIMILAR
TO
KLP
5^
CELLS
............................
55
3.3.7
KLP5-T224D
AFFECTS
THE
OSCILLATORY
MOVEMENT
OF
CHROMOSOMES
DURING
PROMETAPHASE
...........................................................................................................................
58
3.3.8
KLP5-T224D
IS
IMPAIRED
IN
DEPOLYMERIZATION-INDUCING
ACTIVITY,
WHILE
STILL
BEING
ABLE
TO
MOVE
ALONG
MICROTUBULES.
..........................................................................................
60
3.3.9
KLP5-T224D-GFP/KLP6-GFP
SHOWS
PLUS-END
DIRECTED
MOVEMENT
ALONG
ASTRAL
MTS
...........................................................................................................................................65
3.4
D
ISCUSSION
.......................................................................................................................
67
3.4.1
1S
KINESIN-8
(KLP5/KLP6)
A
PHYSIOLOGICAL
SUBSTRATE
OF
AURORA
B/ARKL
?................
68
3.4.2
THE
MODIFICATION
OF
KLP5-T224
RESIDUE
MIGHT
AFFECT
ITS
DEPOLYMERIZATION
ACTIVITY
69
3.5
S
UPPLEMENTAL
DATA
......................................................................................................
73
4
MATERIALS
AND
METHODS
___________________________________
78
4.1
M
ATERIALS
........................................................................................................................
78
4.1.1
YEAST
MEDIA
.................................................................................................................
78
4.1.2
REFERENCES
FOR
S.
POMBE
STRAIN
.................................................................................
79
4.1.3
FISSION
YEAST
STRAINS
USED
IN
CHAPTER
2
...................................................................
80
4.1.4
FISSION
YEAST
STRAINS
USED
IN
CHAPTER
3
...................................................................
81
4.1.5
PLASMIDS
......................................................................................................................
83
4.1.6
ANTIBODIES
..................................................................................................................
84
4.1.7
OLIGONUCLEOTIDES
........................................................................................................
84
4.2
M
ETHODS
..........................................................................................................................
85
4.2.1
CONSTRUCTION
OF
FISSION
YEAST
STRAINS
........................................................................
85
4.2.2
TETRAD
DISSECTION
........................................................................................................
86
4.2.3
GROWTH
TEST
..................................................................................................................
86
IN
4.2.4
YEAST
CULTURE
................................................................................................................
86
4.2.5
CONSTRUCTION
OF
DNA
PLASMIDS
AND
RECOMBINANT
PROTEIN
EXPRESSION
..................
8
7
4.2.6
ARKL
IN
VITRO
KINASE
ASSAY
..........................................................................................
88
4.2.7
PURIFICATION
OF
KLP5/KLP6
FROM
MITOTIC
S.
POMBE
CULTURES
....................................
88
4.2.8
WHOLE
CELL
EXTRACT
PREPARATION
USING
TCA
.............................................................
89
4.2.9
SDS-PA
GE
AND
IMMUNOBLOTTING
...............................................................................90
4.2.10
IMMUNOSTAINING
AND
DNA
STAINING
.........................................................................
90
4.2.11
FLUORESCENCE
MICROSCOPY
.........................................................................................
91
4.2.12
LIVE
CELL
IMAGING
.......................................................................................................
91
5
REFERENCES
------------------------------------------------------------------------------------------------------------------93
CURRICULUM
VITAE....
.................................111
IV
|
| any_adam_object | 1 |
| author | Huang, Yu-Hua 1975- |
| author_GND | (DE-588)142810592 |
| author_facet | Huang, Yu-Hua 1975- |
| author_role | aut |
| author_sort | Huang, Yu-Hua 1975- |
| author_variant | y h h yhh |
| building | Verbundindex |
| bvnumber | BV036965354 |
| ctrlnum | (OCoLC)706976632 (DE-599)DNB1008686581 |
| dewey-full | 571.844742956 |
| dewey-hundreds | 500 - Natural sciences and mathematics |
| dewey-ones | 571 - Physiology & related subjects |
| dewey-raw | 571.844742956 |
| dewey-search | 571.844742956 |
| dewey-sort | 3571.844742956 |
| dewey-tens | 570 - Biology |
| discipline | Biologie |
| format | Thesis Book |
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| indexdate | 2024-07-09T22:51:42Z |
| institution | BVB |
| language | English |
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| physical | IX, 111 S. Ill., graph. Darst. |
| publishDate | 2010 |
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| spelling | Huang, Yu-Hua 1975- Verfasser (DE-588)142810592 aut The fission yeast Aurora kinase possible mechanisms of activation and its role in regulating microtubule dynamics von Yu-Hua Huang 2010 IX, 111 S. Ill., graph. Darst. txt rdacontent n rdamedia nc rdacarrier Tübingen, Univ., Diss., 2010 (DE-588)4113937-9 Hochschulschrift gnd-content DNB Datenaustausch application/pdf http://bvbr.bib-bvb.de:8991/F?func=service&doc_library=BVB01&local_base=BVB01&doc_number=020880166&sequence=000001&line_number=0001&func_code=DB_RECORDS&service_type=MEDIA Inhaltsverzeichnis |
| spellingShingle | Huang, Yu-Hua 1975- The fission yeast Aurora kinase possible mechanisms of activation and its role in regulating microtubule dynamics |
| subject_GND | (DE-588)4113937-9 |
| title | The fission yeast Aurora kinase possible mechanisms of activation and its role in regulating microtubule dynamics |
| title_auth | The fission yeast Aurora kinase possible mechanisms of activation and its role in regulating microtubule dynamics |
| title_exact_search | The fission yeast Aurora kinase possible mechanisms of activation and its role in regulating microtubule dynamics |
| title_full | The fission yeast Aurora kinase possible mechanisms of activation and its role in regulating microtubule dynamics von Yu-Hua Huang |
| title_fullStr | The fission yeast Aurora kinase possible mechanisms of activation and its role in regulating microtubule dynamics von Yu-Hua Huang |
| title_full_unstemmed | The fission yeast Aurora kinase possible mechanisms of activation and its role in regulating microtubule dynamics von Yu-Hua Huang |
| title_short | The fission yeast Aurora kinase |
| title_sort | the fission yeast aurora kinase possible mechanisms of activation and its role in regulating microtubule dynamics |
| title_sub | possible mechanisms of activation and its role in regulating microtubule dynamics |
| topic_facet | Hochschulschrift |
| url | http://bvbr.bib-bvb.de:8991/F?func=service&doc_library=BVB01&local_base=BVB01&doc_number=020880166&sequence=000001&line_number=0001&func_code=DB_RECORDS&service_type=MEDIA |
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