title: Bruton's tyrosine kinase: cell biology, sequence conservation, mutation spectrum, siRNA modifications, and expression profiling.
abstract: Bruton's tyrosine kinase (Btk) is encoded by the gene that when mutated causes the primary immunodeficiency disease X-linked agammaglobulinemia (XLA) in humans and X-linked immunodeficiency (Xid) in mice. Btk is a member of the Tec family of protein tyrosine kinases (PTKs) and plays a vital, but diverse, modulatory role in many cellular processes. Mutations affecting Btk block B-lymphocyte development. Btk is conserved among species, and in this review, we present the sequence of the full-length rat Btk and find it to be analogous to the mouse Btk sequence. We have also analyzed the wealth of information compiled in the mutation database for XLA (BTKbase), representing 554 unique molecular events in 823 families and demonstrate that only selected amino acids are sensitive to replacement (P < 0.001). Although genotype-phenotype correlations have not been established in XLA, based on these findings, we hypothesize that this relationship indeed exists. Using short interfering-RNA technology, we have previously generated active constructs downregulating Btk expression. However, application of recently established guidelines to enhance or decrease the activity was not successful, demonstrating the importance of the primary sequence. We also review the outcome of expression profiling, comparing B lymphocytes from XLA-, Xid-, and Btk-knockout (KO) donors to healthy controls. Finally, in spite of a few genes differing in expression between Xid- and Btk-KO mice, in vivo competition between cells expressing either mutation shows that there is no selective survival advantage of cells carrying one genetic defect over the other. We conclusively demonstrate that for the R28C-missense mutant (Xid), there is no biologically relevant residual activity or any dominant negative effect versus other proteins.
authors.affiliation: Clinical Research Center, Karolinska Institutet, Huddinge, Sweden.
authors.forename: Jessica M
authors.initials: JM
authors.lastname: Lindvall
authors.forename: K Emelie M
authors.initials: KE
authors.lastname: Blomberg
authors.forename: Jouni
authors.initials: J
authors.lastname: Väliaho
authors.forename: Leonardo
authors.initials: L
authors.lastname: Vargas
authors.forename: Juhana E
authors.initials: JE
authors.lastname: Heinonen
authors.forename: Anna
authors.initials: A
authors.lastname: Berglöf
authors.forename: Abdalla J
authors.initials: AJ
authors.lastname: Mohamed
authors.forename: Beston F
authors.initials: BF
authors.lastname: Nore
authors.forename: Mauno
authors.initials: M
authors.lastname: Vihinen
authors.forename: C I Edvard
authors.initials: CI
authors.lastname: Smith
mesh_terms: D000077329:Agammaglobulinaemia Tyrosine Kinase; D000361:Agammaglobulinemia / Q000235:genetics*; D000595:Amino Acid Sequence; D000818:Animals; D017124:Conserved Sequence; D020869:Gene Expression Profiling; D006801:Humans; D007153:Immunologic Deficiency Syndromes / Q000235:genetics*; D051379:Mice; D008969:Molecular Sequence Data; D009154:Mutation; D011505:Protein-Tyrosine Kinases / Q000737:chemistry* / Q000235:genetics* / Q000378:metabolism; D034741:RNA, Small Interfering / Q000235:genetics; D051381:Rats; D016415:Sequence Alignment
chemical_list: D034741:RNA, Small Interfering; D011505:Protein-Tyrosine Kinases; D000077329:Agammaglobulinaemia Tyrosine Kinase; C000625949:BTK protein, human; C000625951:Btk protein, mouse