8 research outputs found
P-Element Homing Is Facilitated by engrailed Polycomb-Group Response Elements in Drosophila melanogaster
P-element vectors are commonly used to make transgenic Drosophila and generally insert in the genome in a nonselective manner. However, when specific fragments of regulatory DNA from a few Drosophila genes are incorporated into P-transposons, they cause the vectors to be inserted near the gene from which the DNA fragment was derived. This is called P-element homing. We mapped the minimal DNA fragment that could mediate homing to the engrailed/invected region of the genome. A 1.6 kb fragment of engrailed regulatory DNA that contains two Polycomb-group response elements (PREs) was sufficient for homing. We made flies that contain a 1.5kb deletion of engrailed DNA (enΔ1.5) in situ, including the PREs and the majority of the fragment that mediates homing. Remarkably, homing still occurs onto the enΔ1. 5 chromosome. In addition to homing to en, P[en] inserts near Polycomb group target genes at an increased frequency compared to P[EPgy2], a vector used to generate 18,214 insertions for the Drosophila gene disruption project. We suggest that homing is mediated by interactions between multiple proteins bound to the homing fragment and proteins bound to multiple areas of the engrailed/invected chromatin domain. Chromatin structure may also play a role in homing
Genome-Wide Tissue-Specific Occupancy of the Hox Protein Ultrabithorax and Hox Cofactor Homothorax in Drosophila
The Hox genes are responsible for generating morphological diversity along the
anterior-posterior axis during animal development. The
Drosophila Hox gene Ultrabithorax
(Ubx), for example, is required for specifying the identity
of the third thoracic (T3) segment of the adult, which includes the dorsal
haltere, an appendage required for flight, and the ventral T3 leg.
Ubx mutants show homeotic transformations of the T3 leg
towards the identity of the T2 leg and the haltere towards the wing. All Hox
genes, including Ubx, encode homeodomain containing
transcription factors, raising the question of what target genes
Ubx regulates to generate these adult structures. To
address this question, we carried out whole genome ChIP-chip studies to identify
all of the Ubx bound regions in the haltere and T3 leg imaginal discs, which are
the precursors to these adult structures. In addition, we used ChIP-chip to
identify the sites bound by the Hox cofactor, Homothorax (Hth). In contrast to
previous ChIP-chip studies carried out in Drosophila embryos,
these binding studies reveal that there is a remarkable amount of tissue- and
transcription factor-specific binding. Analyses of the putative target genes
bound and regulated by these factors suggest that Ubx regulates many downstream
transcription factors and developmental pathways in the haltere and T3 leg.
Finally, we discovered additional DNA sequence motifs that in some cases are
specific for individual data sets, arguing that Ubx and/or Hth work together
with many regionally expressed transcription factors to execute their functions.
Together, these data provide the first whole-genome analysis of the binding
sites and target genes regulated by Ubx to specify the morphologies of the adult
T3 segment of the fly