title: Identification of nuclear protein targets for six leukemogenic tyrosine kinases governed by post-translational regulation.
abstract: Mutated tyrosine kinases are associated with a number of different haematological malignancies including myeloproliferative disorders, lymphoma and acute myeloid leukaemia. The potential commonalities in the action of six of these leukemogenic proteins on nuclear proteins were investigated using systematic proteomic analysis. The effects on over 3600 nuclear proteins and 1500 phosphopeptide sites were relatively quantified in seven isogenic cell lines. The effects of the kinases were diverse although some commonalities were found. Comparison of the nuclear proteomic data with transcriptome data and cytoplasmic proteomic data indicated that the major changes are due to post-translational mechanisms rather than changes in mRNA or protein distribution. Analysis of the promoter regions of genes whose protein levels changed in response to the kinases showed the most common binding site found was that for NFκB whilst other sites such as those for the glucocorticoid receptor were also found. Glucocorticoid receptor levels and phosphorylation were decreased by all 6 PTKs. Whilst Glucocorticoid receptor action can potentiate NFκB action those proteins where genes have NFκB binding sites were in often regulated post-translationally. However all 6 PTKs showed evidence of NFkB pathway modulation via activation via altered IkB and NFKB levels. Validation of a common change was also undertaken with PMS2, a DNA mismatch repair protein. PMS2 nuclear levels were decreased in response to the expression of all 6 kinases, with no concomitant change in mRNA level or cytosolic protein level. Response to thioguanine, that requires the mismatch repair pathway, was modulated by all 6 oncogenic kinases. In summary common targets for 6 oncogenic PTKs have been found that are regulated by post-translational mechanisms. They represent potential new avenues for therapies but also demonstrate the post-translational regulation is a key target of leukaemogenic kinases.
authors: Stem Cell and Leukaemia Proteomics Laboratory, Manchester Academic Health Science Centre, The University of Manchester, Manchester, United Kingdom.
authors: Andrew
authors: A
authors: Pierce
authors: Andrew
authors: A
authors: Williamson
authors: Ewa
authors: E
authors: Jaworska
authors: John R
authors: JR
authors: Griffiths
authors: Sam
authors: S
authors: Taylor
authors: Michael
authors: M
authors: Walker
authors: Mark
authors: M
authors: Aspinall-O'Dea
authors: Mark Aspinall
authors: MA
authors: O'Dea
authors: Elaine
authors: E
authors: Spooncer
authors: Richard D
authors: RD
authors: Unwin
authors: Toryn
authors: T
authors: Poolman
authors: David
authors: D
authors: Ray
authors: Anthony D
authors: AD
authors: Whetton
mesh_terms: D000818:Animals; D002460:Cell Line; D013058:Mass Spectrometry; D051379:Mice; D009687:Nuclear Proteins / Q000378:metabolism*; D011499:Protein Processing, Post-Translational; D011505:Protein-Tyrosine Kinases / Q000378:metabolism*; D040901:Proteomics; D011965:Receptors, Glucocorticoid / Q000378:metabolism; D015398:Signal Transduction
chemical_list: D009687:Nuclear Proteins; D011965:Receptors, Glucocorticoid; D011505:Protein-Tyrosine Kinases