GAGA factor isoforms have distinct but overlapping functions in vivo.
The Drosophila melanogaster GAGA factor (encoded by the Trithorax-like [Trl] gene) is required for correct chromatin architecture at diverse chromosomal sites. The Trl gene encodes two alternatively spliced isoforms of the GAGA factor (GAGA-519 and GAGA-581) that are identical except for the length and sequence of the C-terminal glutamine-rich (Q) domain. In vitro and tissue culture experiments failed to find any functional difference between the two isoforms. We made a set of transgenes that constitutively express cDNAs coding for either of the isoforms with the goal of elucidating their roles in vivo. Phenotypic analysis of the transgenes in Trl mutant background led us to the conclusion that GAGA-519 and GAGA-581 perform different, albeit largely overlapping, functions. We also expressed a fusion protein with LacZ disrupting the Q domain of GAGA-519. This LacZ fusion protein compensated for the loss of wild-type GAGA factor to a surprisingly large extent. This suggests that the Q domain either is not required for the essential functions performed by the GAGA protein or is exclusively used for tetramer formation. These results are inconsistent with a major role of the Q domain in chromatin remodeling or transcriptional activation. We also found that GAGA-LacZ was able to associate with sites not normally occupied by the GAGA factor, pointing to a role of the Q domain in binding site choice in vivo.
Greenberg
A J
AJ
Dept. of Molecular Biology, Princeton University, Princeton, NJ 08544, USA.
Schedl
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D000818:Animals; D030801:Animals, Genetically Modified / Q000378:metabolism; D001665:Binding Sites; D015153:Blotting, Western; D002467:Cell Nucleus / Q000378:metabolism; D002843:Chromatin / Q000378:metabolism; D003433:Crosses, Genetic; D018076:DNA, Complementary / Q000378:metabolism; D004268:DNA-Binding Proteins; D004330:Drosophila; D029721:Drosophila Proteins; D004331:Drosophila melanogaster; D018398:Homeodomain Proteins / Q000737:chemistry* / Q000235:genetics / Q000502:physiology*; D007763:Lac Operon; D008856:Microscopy, Fluorescence; D008938:Mitosis; D009154:Mutation; D010641:Phenotype; D020033:Protein Isoforms; D017434:Protein Structure, Tertiary; D014157:Transcription Factors / Q000737:chemistry* / Q000235:genetics / Q000502:physiology*; D014158:Transcription, Genetic; D019076:Transgenes; D001616:beta-Galactosidase / Q000378:metabolism
D002843:Chromatin; D018076:DNA, Complementary; D004268:DNA-Binding Proteins; D029721:Drosophila Proteins; D018398:Homeodomain Proteins; D020033:Protein Isoforms; D014157:Transcription Factors; C063460:Trl protein, Drosophila; D001616:beta-Galactosidase