Analysis of chromatin structure of genes silenced by heterochromatin in trans.
While heterochromatic gene silencing in cis is often accompanied by nucleosomal compaction, characteristic histone modifications, and recruitment of heterochromatin proteins, little is known concerning genes silenced by heterochromatin in trans. An insertion of heterochromatic satellite DNA in the euchromatic brown (bw) gene of Drosophila melanogaster results in bwDominant (bwD), which can inactivate loci on the homolog by relocation near the centric heterochromatin (trans-inactivation). Nucleosomal compaction was found to accompany trans-inactivation, but stereotypical heterochromatic histone modifications were mostly absent on silenced reporter genes. HP1 was enriched on trans-inactivated reporter constructs and this enrichment was more pronounced on adult chromatin than on larval chromatin. Interestingly, this HP1 enrichment in trans was unaccompanied by an increase in the 2MeH3K9 mark, which is generally thought to be the docking site for HP1 in heterochromatin. However, a substantial increase in the 2MeH3K9 mark was found on or near the bwD satellite insertion in cis, but did not spread further. These observations suggest that the interaction of HP1 with chromatin in cis is fundamentally different from that in trans. Our molecular data agree well with the differential phenotypic effect on bwD trans-inactivation of various genes known to be involved in histone modification and cis gene silencing.
Nisha
Parul
P
Department of Biological Sciences, Carnegie Mellon University, Pittsburgh, PA 15213, USA.
Plank
Jennifer L
JL
Csink
Amy K
AK
D018528:ATP-Binding Cassette Transporters / Q000235:genetics*; D000818:Animals; D047369:Chromatin Immunoprecipitation; D002868:Chromosomal Proteins, Non-Histone / Q000235:genetics*; D017931:DNA Primers / Q000235:genetics; D029721:Drosophila Proteins / Q000235:genetics*; D004331:Drosophila melanogaster / Q000235:genetics*; D020868:Gene Silencing / Q000502:physiology*; D006570:Heterochromatin / Q000235:genetics* / Q000502:physiology; D009707:Nucleosomes / Q000378:metabolism; D015183:Restriction Mapping
D018528:ATP-Binding Cassette Transporters; D002868:Chromosomal Proteins, Non-Histone; D017931:DNA Primers; D029721:Drosophila Proteins; D006570:Heterochromatin; D009707:Nucleosomes; C059253:bw protein, Drosophila; C452193:heterochromatin protein 1, Drosophila