Abstract
Purpose:
Bruton's tyrosine kinase is crucial for B-lymphocyte development. By the use of gene expression profiling, we have identified four expressed sequence tags among 38 potential Btk target genes, which have now been characterised.
Methods:
Bioinformatics tools including data mining of additional unpublished gene expression profiles, sequence verification of PCR products and qualitative RT-PCR were used. Stimulations targeting the B-cell receptor and the protein kinase C were used to activate whole B-cell splenocytes.
Results:
Target genes were characterised as Lim domain only 7 (Lmo7); Myosin1e (Myo1e); SAM and SH3 domain containing 1 (Sash1); and Mucolipin2 (Mcoln2). Expression was found in cell lines of different origin and developmental stages as well as in whole B-cell splenocytes and Transitional type 1 (T1) splenic B-cells from wild type and Btk-defective mice, respectively. By the use of semi-quantitative RT-PCR we found Sash1 not to be expressed in the investigated haematopoietic cell lines, while transcripts were found in whole splenic B-cells from both wild type and Btk-defective mice, whereas Lmo7, Myo1e, and Mcoln2 were expressed in both B-cell lines and primary B-lymphocytes. Except for Lmo7, the transcript level was similarly affected by stimulation in control and Btk-defective cells.
Publication types
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Research Support, Non-U.S. Gov't
MeSH terms
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Agammaglobulinaemia Tyrosine Kinase
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Animals
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B-Lymphocytes / metabolism*
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Cation Transport Proteins / biosynthesis*
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Cation Transport Proteins / chemistry
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Cation Transport Proteins / genetics
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Cell Line, Tumor
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Computational Biology
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Gene Expression Profiling
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Homeodomain Proteins / biosynthesis
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Homeodomain Proteins / chemistry
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Homeodomain Proteins / genetics*
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LIM Domain Proteins
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Membrane Proteins / biosynthesis*
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Membrane Proteins / chemistry
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Membrane Proteins / genetics
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Mice
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Myosin Type I
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Myosins / biosynthesis*
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Myosins / chemistry
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Myosins / genetics
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NIH 3T3 Cells
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Protein-Tyrosine Kinases / deficiency*
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Protein-Tyrosine Kinases / genetics*
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RNA, Messenger / metabolism
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TRPM Cation Channels
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Transcription Factors / biosynthesis
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Transcription Factors / chemistry
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Transcription Factors / genetics*
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Transient Receptor Potential Channels
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Tumor Suppressor Proteins / biosynthesis*
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Tumor Suppressor Proteins / chemistry
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Tumor Suppressor Proteins / genetics
Substances
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Cation Transport Proteins
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Homeodomain Proteins
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LIM Domain Proteins
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Lmo7 protein, mouse
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Mcoln2 protein, mouse
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Membrane Proteins
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RNA, Messenger
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Sash1 protein, mouse
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TRPM Cation Channels
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Transcription Factors
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Transient Receptor Potential Channels
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Tumor Suppressor Proteins
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Protein-Tyrosine Kinases
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Agammaglobulinaemia Tyrosine Kinase
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Btk protein, mouse
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Myo1e protein, mouse
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Myosin Type I
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Myosins