Regulation of naturally occurring HIV-1 LTRs by MAPK signaling cascades:
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| Format: | Book |
| Language: | English |
| Published: |
2000
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| Subjects: | |
| Online Access: | Inhaltsverzeichnis |
| Item Description: | Zsfassung in dt. Sprache. - Würzburg, Univ., Diss., 2000 |
| Physical Description: | III, 91 Bl. Ill., graph. Darst. |
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| 245 | 1 | 0 | |a Regulation of naturally occurring HIV-1 LTRs by MAPK signaling cascades |c Peifeng Chen |
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TABLE OF CONTENTS
TABLE OF CONTENTS
ABSTRACT_ /
ZVSVMMENFASSVNG
. 2
INTRODUCTION
_ 3
LI. GENERAL BACKGROUND OF HIV-1_3
1.1.1. HIV-1 INFECTION AND PATHOGENESIS.3
1-1.2. TRANSCRIPTIONAL REGULATION OF HIV-1 PROMOTERS. 3
1-1.3. HIV-1 LTR MUTATIONS OR INSERTIONS MAY IMPACT THE COURSE OF HIV-1
DISEASE.6
1-2. MAPK SIGNALING PATHWAYS_7
1-2.1. MITOGEN-ACTIVATED RAF/MEK/ERK SIGNALING CASCADE.8
1-2.2. STRESS-INDUCED SIGNALING CASCADES: THE SAPK/JNK AND P38
PATHWAYS.10
1-2.3. REGULATION OF THE MAPK SIGNALING PATHWAYS BY SCAFFOLD PROTEINS.11
1-2.4. TRANSCRIPTIONAL REGULATION BY THE MAPK SIGNALING PATHWAYS.12
1-2.4.1. REGULATION OF TRANSCRIPTION FACTOR AP-1 ACTIVITY BY MAPKS.12
I-2.4.2. REGULATION OF TRANSCRIPTION FACTOR
NF-KB BY MAPKS.14
1-3. T CELL AS A MODEL SYSTEM FOR THE STUDY OF HIV-1 TRANSCRIPTIONAL
REGULATION . 15
1-3.1. T CEL) ACTIVATION.15
1-3.2. T CELL ACTIVATION AND MAPK SIGNALING PATHWAYS.16
1-3.3. CD4* T CELL ACTIVATION IS IMPORTANT FOR THE HIV-1 GENE
REGULATION.17
I- 4. DESIGN AND STRATEGIES OF THE WORK.
.--.17
RESULTS
.19
II- L. NATURALLY OCCURRING HIV-1 LTRS DISPLAY A WIDE-RANGE OF BASAL AND
TAT-MEDIATED
TRANSCRIPTIONAL ACTIVITY.YYMM.M.~.~.-19
II-2. TRANSACTIVATION OF NATURALLY OCCURRING HIV-1 LTRS BY MITOGENIC AND
STRESS INDUCED
SIGNALING PATHWAYS. 20
II-3. DOMINANT NEGATIVE KINHSES AND INHIBITORS OF MAPK SIGNALING
PATHWAYS REDUCED
TRANSCRIPTIONAL ACTIVITY OF THE NATURALLY OCCURRING HIV-1 LTR
BT94T-BL.21
B-3.1.TPA- AND TPA/ION-INDUCED RAF/MEK/ERK SIGNALING PATHWAY IS
PARTIALLY INHIBITED BY ERK-B3,
ERK-C3 AND RAF-C4HA, AND MEK INHIBITOR U0126.22
D-3.2. MKK6(ALA), SPRK(K A) ARE NOT SUFFICIENT TO INHIBIT TPA AND
TPA/ION INDUCED TRANSACTIVATION OF
HIV-1 PROMOTERS.24
N-4. ACTIVE KINASES FROM THE ERK-, SAPK/JNK-, AND P38-ACTIVATING
PATHWAYS TRANSACTIVATE
NATURALLY OCCURRING HIV-1 LTRS.-.25
D-4.1. RAF-BXB-CAAX TRANSACTIVATE NATURALLY OCCURRING HIV-1 LTRS.25
D-4.2. MKK6CEE) TRANSACTIVATES NATURALLY OCCURRING HIV-LTRS VIA P38
PATHWAY.26
B-4.3. SPRK-INDUCED SAPK/JNK SIGNALING PATHWAY IS INVOLVED IN THE
TRANSCRIPTIONAL REGULATION OF THE
NATURALLY OCCURRING HIV-1 LTR VARIANTS.27
N-5. SPRK AND
NF-KB BINDING SITES OF HIV-1 LTR-30
H-5.1. SPRK TRANSACTIVATE HIV-1 NF-KB.30
B-5.2. IKKPKD AND JBD SYNERGISTICALLY INHIBIT SPRK-INDUCED
TRANSACTIVATION OF NATURALLY OCCURRING
HIV-1 PROMOTER BT94T-B 1.30
I
BIBLIOGRAFISCHE INFORMATIONEN
HTTP://D-NB.INFO/959865276
TABLE OF CONTENTS
N-6.
AP-L FACTOR* BIND TO THE MFNLP SEQUENCE
AT
THE HIV-1 PROMOTER BT94T-BL IN T CELLS
.32
D-6.1. FIVE PROTEIN COMPLEXES BIND TO THE BT94T-BL SPECIFIC MFNLP
SEQUENCE.32
D-6.2. THE 3* HALF OF THE MFNLP SEQUENCE IS CRUCIAL FOR THE PROTEIN-DNA
INTERACTION.34
0-6.3. THE SECOND PROTEIN COMPLEX CONTAINS AP-L FACTORS.34
H-7. MFNLP SEQUENCE SHARES SIMILAR FACTORS WITH SERUM RESPONSIVE
ELEMENTS OF THE C-FOS
PMNNTAR~--- --.-.34
D-8. POINT MUTATIONS INTRODUCED TO THE MFNLP SEQUENCE HAVE AN EFFECT ON
THE BINDING OF
AP-L FACTOR-CONTAINING COMPLEX AND TRAMMRRIWHNN ., ,YYYY
YYYYYYYYYYYYYY
. 38
D-8.1. POINT MUTATIONS INTRODUCED TO THE MFNLP SEQUENCE ABOLISHED
BINDING OF THE AP-L FACTOR-CONTAINING
COMPLEXES.
38
0-8.2. POINT MUTATIONS INTRODUCED TO THE MFNLP SEQUENCE OF BT94T-B 1
PROMOTER ENHANCE TPA-, AND
TP A/ION - INDUCED TRANSACTIVATION.40
0-8.3. POINT MUTATIONS INTRODUCED TO THE MFNLP SEQUENCE OF BT94T-B 1
PROMOTER REDUCE SPRK-INDUCED
TRANSACTIVATION.
40
DISCUSSION
.
42
IH-1. NATURALLY OCCURRING HIV-1 LTRS DISPLAY A WIDE-RANGE OF BASAL AND
TAT-MEDIATED
TRANSCRIPTIONAL ARRIVISTE*.,.::.T.-T-TIRTTRRYY.42
III-2. T CELL ACTIVATION AND ACTIVATION OF MAPK KINASES LEAD TO THE
TRANSACTIVATION OF
NATURALLY OCCURRING HIV-1 LTRS.-.--.,.
43
HI-3. MFNLP INSERTION IS IMPORTANT FOR THE SPRK MEDIATED
TRANS-ACTIVATION OF PROMOTER
BT94T-B1.-.
47
III-4. CONCLUDING REMARK*--.
.
T-49
MATERIALS AND METHODS
.5/
IV-1. MATERIALS.
IV-1.1. INSTRUMENTS.
IV-1.2. REAGENTS AND GENERAL MATERIALS.
IV-1.4. CELL CULTURE MATERIALS.
IV-1.5. ENZYMES.*.
IV-1.6. ANTIBODY CONJUGATES.
IV-1.7. KITS.
IV-1.8. PLASMIDS.
IV-1.9. OLIGONUCLEOTIDES.
IV-1.10. BACTERIA STRAIN AND CELL CULTURE
IV-1.11. MEDIA.
LB (LUNA-BERTANI) MEDIUM.
GC (CIRCLE-GROW) MEDIUM.
2X TY MEDIUM.
IV-1.12. SOLUTIONS.
IV-2. METHODS.
IV-2.1. WORKING WITH BACTERIA.
IV-2.1.1. BACTERIAL CULTURES.
IV-2.1.2. PREPARATION OF COMPETENT CELLS (CACL2 METHOD),
IV-2.1.3. TRANSFORMATION OF COMPETENT BACTERIA.
IV-2.2. WORKING WITH DNAS.
51
51
51
52
53
53
53
53
54
55
55
55
55
55
55
59
59
59
59
60
60
11
TABIC OF CONTENTS
IV-2.2.1. ELEC
TROPHORESIS OF DNA ON AGAROSE GEL.60
IV-2.2.2. ISOLATION OF PLASMID DNA FROM AGAROSE (QIAEX D AGAROSE GEL
EXTRACTION PROTOCOL).60
TV-2.2.3. PURIFICATION OF PLASMID DNA (QLAQUICK PCR PURIFICATION KIT).61
TV-22.4.
LIGATION OF DNA FRAGMENTS.61
IV-2.2.5. MINI-PREPARATION ERF PLASMID DNA.61
IV-2.2.6. MAXI-PREPARATION OF PLASMID DNA.62
IV-2.2.7. MEASUREMENT OF DNA CONCENTRATION.62
IV-2.2.8. DNA SEQUENCING (SANGER DIDEOXY METHOD).62
IV-2.2.9. PCR SITE DIRECTED MUTAGENESIS (QUIKCHANGE
YY
SITE-DIRECTED MUTAGENESIS).63
IV-2.3. WORKING WITH PROTEINS.64
IV-2.3.1. IMMUNOPRECIPITATION.64
IV-2.3.2.
IN VITRO KINASE ASSAY. 65
IV-2.3.3. MEASUREMENT OF PROTEIN CONCENTRATION (BIO-RAD PROTEIN
ASSAY).65
FV-2.3.4. SODIUM DODECYL SULFATE POLYACRYLAMIDE GEL ELECTROPHORESIS (SDS
PAGE).65
IV-2.3.5. IMMUNOBOTTING.66
IV-2.4. WORKING WITH CELL CULTURES. 66
IV-2.4.1. MAINTENANCE OF CELL CULTURE.66
IV-2.4.2. TRANSIENT TRANSFECTION ASSAY (DMWE
YY
-C LIPOSOME METHOD).67
IV-2.4.3, LUCIFERASE REPORTER GENE ASSAY.67
IV-2.4.4, FI-GALACTOSIDASE ASSAY.68
IV-2.5. PROTEIN-DNA INTERACTION.68
IV-2.5.1. PREPARATION OF CYTOPLASMIC AND NUCLEAR EXTRACTS.68
IV-2.5.2. ANNEALING REACTION OF OLIGONUCLEOTIDES.68
IV-2.5.3. RANDOM LABELING OF OLIGONUCLEOTIDES BY KLENOW METHOD.68
IV-2.5.4. ELECTROPHORETIC GEL MOBILITY SHIFT ASSAY (EMSA).69
REFERENCES
APPENDIX
.
ABBREVIATIONS.
70
88
88
CURRICULUM VITAE.
_90
IN |
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| indexdate | 2024-10-09T18:10:53Z |
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| spelling | Chen, Peifeng 1968- Verfasser (DE-588)122374258 aut Regulation of naturally occurring HIV-1 LTRs by MAPK signaling cascades Peifeng Chen 2000 III, 91 Bl. Ill., graph. Darst. txt rdacontent n rdamedia nc rdacarrier Zsfassung in dt. Sprache. - Würzburg, Univ., Diss., 2000 Promotor Genetik (DE-588)4263977-3 gnd rswk-swf HIV (DE-588)4200792-6 gnd rswk-swf Replikation (DE-588)4153174-7 gnd rswk-swf Genmutation (DE-588)4156638-5 gnd rswk-swf (DE-588)4113937-9 Hochschulschrift gnd-content HIV (DE-588)4200792-6 s Promotor Genetik (DE-588)4263977-3 s Genmutation (DE-588)4156638-5 s Replikation (DE-588)4153174-7 s DE-604 DNB Datenaustausch application/pdf http://bvbr.bib-bvb.de:8991/F?func=service&doc_library=BVB01&local_base=BVB01&doc_number=009009189&sequence=000001&line_number=0001&func_code=DB_RECORDS&service_type=MEDIA Inhaltsverzeichnis |
| spellingShingle | Chen, Peifeng 1968- Regulation of naturally occurring HIV-1 LTRs by MAPK signaling cascades Promotor Genetik (DE-588)4263977-3 gnd HIV (DE-588)4200792-6 gnd Replikation (DE-588)4153174-7 gnd Genmutation (DE-588)4156638-5 gnd |
| subject_GND | (DE-588)4263977-3 (DE-588)4200792-6 (DE-588)4153174-7 (DE-588)4156638-5 (DE-588)4113937-9 |
| title | Regulation of naturally occurring HIV-1 LTRs by MAPK signaling cascades |
| title_auth | Regulation of naturally occurring HIV-1 LTRs by MAPK signaling cascades |
| title_exact_search | Regulation of naturally occurring HIV-1 LTRs by MAPK signaling cascades |
| title_full | Regulation of naturally occurring HIV-1 LTRs by MAPK signaling cascades Peifeng Chen |
| title_fullStr | Regulation of naturally occurring HIV-1 LTRs by MAPK signaling cascades Peifeng Chen |
| title_full_unstemmed | Regulation of naturally occurring HIV-1 LTRs by MAPK signaling cascades Peifeng Chen |
| title_short | Regulation of naturally occurring HIV-1 LTRs by MAPK signaling cascades |
| title_sort | regulation of naturally occurring hiv 1 ltrs by mapk signaling cascades |
| topic | Promotor Genetik (DE-588)4263977-3 gnd HIV (DE-588)4200792-6 gnd Replikation (DE-588)4153174-7 gnd Genmutation (DE-588)4156638-5 gnd |
| topic_facet | Promotor Genetik HIV Replikation Genmutation Hochschulschrift |
| url | http://bvbr.bib-bvb.de:8991/F?func=service&doc_library=BVB01&local_base=BVB01&doc_number=009009189&sequence=000001&line_number=0001&func_code=DB_RECORDS&service_type=MEDIA |
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