The glutamine-rich domain of the Drosophila GAGA factor is necessary for amyloid fibre formation in vitro, but not for chromatin remodelling.
The Drosophila GAGA factor binds specifically to the sequence GAGAG, and synergises with nucleosome remodelling factor to remodel chromatin in vitro. It consists of an N-terminal domain (POZ/BTB) which mediates protein-protein interactions, a central region which contains the DNA-binding domain, and a C-terminal glutamine-rich region. It is shown that the glutamine-rich region is responsible for the formation of fibres in vitro which, on the basis of their tinctorial properties and CD spectra, may be classified as amyloid fibres. A large structural change, probably resulting in beta-sheet structure, is observed upon fibre formation. Mutants containing the central region, either alone or together with the glutamine-rich region, are largely lacking in secondary structure but they bind specifically to the cognate DNA and are able to remodel chromatin in vitro. Consequently, neither the N-terminal domain nor the C-terminal glutamine-rich regions of the GAGA factor are necessary for chromatin remodelling in vitro.
Agianian
B
B
Structural Biology Programme, European Molecular Biology Laboratory, Meyerhofstrasse 1 D-69117, Heidelgberg, Germany.
Leonard
K
K
Bonte
E
E
Van der Zandt
H
H
Becker
P B
PB
Tucker
P A
PA
D000682:Amyloid / Q000737:chemistry / Q000502:physiology*; D000818:Animals; D052160:Benzothiazoles; D001665:Binding Sites; D001718:Birefringence; D002843:Chromatin / Q000502:physiology*; D003224:Congo Red; D004268:DNA-Binding Proteins / Q000235:genetics / Q000502:physiology*; D029721:Drosophila Proteins; D004331:Drosophila melanogaster; D005456:Fluorescent Dyes; D005973:Glutamine / Q000235:genetics / Q000502:physiology*; D018398:Homeodomain Proteins / Q000235:genetics / Q000502:physiology*; D016296:Mutagenesis; D017433:Protein Structure, Secondary; D011993:Recombinant Fusion Proteins / Q000235:genetics; D013329:Structure-Activity Relationship; D013844:Thiazoles; D015534:Trans-Activators / Q000737:chemistry; D014157:Transcription Factors / Q000235:genetics / Q000502:physiology*
D000682:Amyloid; D052160:Benzothiazoles; D002843:Chromatin; D004268:DNA-Binding Proteins; D029721:Drosophila Proteins; D005456:Fluorescent Dyes; D018398:Homeodomain Proteins; D011993:Recombinant Fusion Proteins; D013844:Thiazoles; D015534:Trans-Activators; D014157:Transcription Factors; C063460:Trl protein, Drosophila; D005973:Glutamine; C009462:thioflavin T; D003224:Congo Red