The Drosophila microtubule associated protein Futsch is phosphorylated by Shaggy/Zeste-white 3 at an homologous GSK3beta phosphorylation site in MAP1B.
The Drosophila homologue of the microtubule associated protein MAP1B is encoded by the futsch locus. The deduced protein Futsch is about twice the size of MAP1B and shows high homology in the N- and C-terminal domains. The central part of Futsch is characterized by a highly repetitive structure based on a 37 amino acid motif. Futsch, like MAP1B, colocalizes with microtubules and is necessary for the organization of the microtubule cytoskeleton during axonal growth and synaptogenesis. To further analyze the functional relevance of Futsch as a MAP1B-like protein, we performed a molecular analysis of the conserved protein domains. Using a number of antisera, we show that, unlike the MAP1B polyprotein, which is cleaved to generate a heavy and light chain, Futsch is expressed as a single protein. The function of MAP1B is in part regulated by phosphorylation mediated by kinases that include casein kinase 2 and glycogen synthase kinase 3beta (GSK3beta). We show here that at least one GSK3beta phosphorylation site of MAP1B is conserved in Futsch and that this site can be phosphorylated by GSK3beta and its Drosophila homologue, Shaggy/Zeste-white 3. To test the functional relevance of these findings we generated a number of minigenes and assayed their ability to rescue the phenotype of futsch mutants. Our data highlight some differences between MAP1B and Futsch but demonstrate that important structural and functional aspects are conserved between fly and vertebrate members of this protein family.
Gögel
Stefanie
S
The MRC Centre for Developmental Neurobiology, New Hunt's House, Guy's Campus, King's College London, London SE1 1UL, UK.
Wakefield
Sarah
S
Tear
Guy
G
Klämbt
Christian
C
Gordon-Weeks
Phillip R
PR
D000595:Amino Acid Sequence; D000818:Animals; D030801:Animals, Genetically Modified; D000918:Antibody Specificity; D017124:Conserved Sequence; D004330:Drosophila; D029721:Drosophila Proteins / Q000737:chemistry / Q000235:genetics* / Q000276:immunology / Q000378:metabolism*; D000939:Epitopes; D038362:Glycogen Synthase Kinase 3 / Q000737:chemistry / Q000235:genetics* / Q000276:immunology / Q000378:metabolism*; D000071679:Glycogen Synthase Kinase 3 beta; D008322:Mammals; D008869:Microtubule-Associated Proteins / Q000737:chemistry / Q000235:genetics* / Q000276:immunology / Q000378:metabolism*; D008870:Microtubules / Q000378:metabolism; D009154:Mutation; D009414:Nerve Growth Factors / Q000737:chemistry / Q000235:genetics* / Q000276:immunology / Q000378:metabolism*; D009469:Neuromuscular Junction / Q000502:physiology; D010766:Phosphorylation; D017434:Protein Structure, Tertiary; D013045:Species Specificity
D029721:Drosophila Proteins; D000939:Epitopes; D008869:Microtubule-Associated Proteins; D009414:Nerve Growth Factors; C409635:futsch protein, Drosophila; C085284:microtubule-associated protein 1B; D000071679:Glycogen Synthase Kinase 3 beta; C080335:Sgg protein, Drosophila; D038362:Glycogen Synthase Kinase 3