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@PHDTHESIS{Widowati:730419,
      author       = {Widowati, Esti Wahyu},
      othercontributors = {Becker, Walter and Claßen-Linke, Irmgard},
      title        = {{F}unctional characterization of {DYRK}1{A} point mutants
                      related to human monogenic disorders},
      school       = {RWTH Aachen University},
      type         = {Dissertation},
      address      = {Aachen},
      reportid     = {RWTH-2018-226842},
      pages        = {1 Online-Ressource : Illustrationen},
      year         = {2018},
      note         = {Veröffentlicht auf dem Publikationsserver der RWTH Aachen
                      University; Dissertation, RWTH Aachen University, 2018,
                      Kumulative Dissertation},
      abstract     = {Functional characterization of DYRK1A point mutants related
                      to human monogenic disorders(Esti Wahyu Widowati)DYRK1A is a
                      member of DYRK (dual-specificity tyrosine (Y)
                      phosphorylation-regulated kinase) family of protein kinases
                      which share conserved structure of the DH (DYRK-homology)
                      box and kinase domain, but differ in their N- and C-terminal
                      sequences. DYRK1A achieves full catalytic activity by
                      tyrosine autophosphorylation which takes place as a one-time
                      event during or immediately after translation. Several
                      truncation mutations, microdeletions and missense variants
                      result in the neurodevelopmental syndrome termed mental
                      retardation autosomal dominant 7 (MRD7). The phenotype of
                      MRD7 includes microcephaly, intellectual disability,
                      epileptic seizures, autism spectrum disorder and language
                      delay and is caused by haploinsufficiency of DYRK1A. This
                      study aims to address missense mutations in kinase domain
                      and provide a functional analysis of those mutants. To
                      characterize the disease-causing missense variants that
                      affect the catalytic domain of DYRK1A, we used a mammalian
                      expression system. Four of the substitutions eliminated
                      tyrosine autophosphorylation (L245R, F308V, S311F, S346P),
                      indicating that these variants lacked kinase activity.
                      Tyrosine phosphorylation of DYRK1A-L295F in mammalian cells
                      was comparable to wild type although this mutant showed
                      lower catalytic activity and reduced thermodynamic stability
                      in cellular thermal assays. One variant (DYRK1A-T588N) with
                      a mutation outside the catalytic domain did not differ from
                      wild type DYRK1A in tyrosine autophosphorylation, catalytic
                      activity or cellular localization. In addition, we
                      investigated two DYRK1A mutants (D138P and K150C) located in
                      the DH-box which correspond to missense variants in the
                      related DYRK1B kinase that are associated with a familial
                      form of metabolic syndrome (AOMS3). The DH-box contributes
                      to the conformational stability of catalytic domain in
                      DYRK1A. Expression in HeLa cells showed that there is no
                      significant difference between DYRK1A-D138P and K150C
                      regarding tyrosine autophosphorylation or catalytic
                      activity. However, reduced tyrosine phosphorylation was
                      observed in both DYRK1A variants when expressed in a
                      bacterial cell free in vitro translation system. In summary,
                      these studies suggest that 1) pathogenic missense variants
                      in the catalytic domain of DYRK1A impair enzymatic function
                      by affecting catalytic amino acid residues or by
                      compromising the structural integrity of the kinase domain
                      and 2) D138 and K150 participate in the maturation of DYRK1A
                      albeit these mutations are compensated under physiological
                      conditions.},
      cin          = {528500-2},
      ddc          = {620},
      cid          = {$I:(DE-82)528500-2_20140620$},
      typ          = {PUB:(DE-HGF)11},
      doi          = {10.18154/RWTH-2018-226842},
      url          = {https://publications.rwth-aachen.de/record/730419},
}