V-Myb proteins and their oncogenic potential : a study on how two point mutations affect the interaction of v-Myb with other proteins / Beeke Wienert.

Format:

Book

Author/Creator:

Wienert, Beeke.

Language:

English

Subjects (All):

Protein-protein interactions.

Oncogenic viruses.

Proteins.

Physical Description:

1 online resource (118 p.)

Edition:

1st ed.

Place of Publication:

Hamburg : Diplomica Verlag, 2012.

Language Note:

English

Summary:

The oncogene v-myb of the retroviruses AMV (avian myeloblastosis virus) and E26 (avian leukaemia virus) encodes a transcription factor (v-Myb) which is a truncated homolog of its cellular progenitor c-Myb. c-Myb plays an essential role in the development of haematopoietic cells and is known to be a regulator for many target genes. v-Myb AMV is responsible for the transformation of myelomonocytic cells and for arresting them in an immature stage, presumably because of a deregulation of the expression of specific target genes. In addition to the truncation of the coding region, a number of amino

Contents:

v-Myb proteins and their oncogenic potential; Table of contents; Abbreviations; 1 Abstract; 2 Introduction; 2.1 The myb gene family of transcription factors; 2.2 The haematopoeietic system; 2.3 The mim-1 gene as a model for gene regulation by v-Myb; 2.4 Aim of the study; 3 Material; 3.1 Chemicals; 3.2 Kits; 3.3 Devices and instruments; 3.4 Enzymes; 3.5 Antibodies; 3.6 Plasmids; 3.7 Oligonucleotides; 3.8 Bacterial strains; 3.9 Media and agar plates; 3.10 Cell culture and materials; 3.11 Cell lines; 3.12 Cell culture media; 3.13 Buffers and solutions; 4 Methods

4.1 Preparation of competent bacteria4.2 Transformation of competent bacteria; 4.3 Plasmid DNA isolation; 4.4 Quantification of nucleic acids; 4.5 Modification of DNA by enzymes; 4.6 Agarose gel electrophoresis; 4.7 DNA fragment extraction; 4.8 Ligation; 4.9 Polymerase chain reaction (PCR); 4.10 Passage and cultivation of cells; 4.11 Transient transfection by calcium phosphate co-precipitation; 4.12 Transient transfection by lipofection with MetafectenePro; 4.13 Bacterial GST-fusion protein expression and purification; 4.14 Protein extraction from eukaryotic cells; 4.15 SDS PAGE

4.16 Gel staining4.17 Western blot and immuno detection; 4.18 GST pull-down assay; 4.19 GFP/ YFP trap; 4.20 Co-immunoprecipitation; 4.21 Reporter gene assay; 5 Results; 5.1 Introduction of different constructs of v-Myb; 5.2 Analysis of the interaction of the hydrophobic region of v-Myb with unidentified binding partners; 5.3 Analysis of the interaction of v-Myb with GRP78; 5.4 Analysis of the interaction between C/EBPß and the hydrophobic region of v-Myb; 5.5 Analysis of the interaction of PRMT4/CARM1 with v-Myb; 6 Discussion

6.1 Endogenous pull-down experiments detected potential interaction partners of the hydrophobic region of v-Myb6.2 GRP78va interacts with both mutants of v-Myb; 6.3 C/EBPß interacts with the hydrophobic region of v-Myb; 6.4 PRMT4 as a newly identified interaction partner; 6.5 Future perspectives; 7 Appendix; 7.1 Table of figures; 7.2 References; 7.3 Clone charts

Notes:

Cover title.

Includes bibliographical references.

ISBN:

9783842832916

3842832915

OCLC:

854977290