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Series GSE3055

Query DataSets for GSE3055

Status

Public on Aug 05, 2005

Title

Sex-specific role of Drosophila HP1 in regulating chromatin structure and gene transcription

Organism

Drosophila melanogaster

Experiment type

Expression profiling by array

Summary

Drosophila heterochromatin protein 1- HP11 is believed to be involved in active transcription, transcriptional gene silencing, and the formation of heterochromatin2-7. However, little is known about the function of HP1 during development. Using a Gal4-induced RNA interference system, we show that conditional depletion of HP1 in transgenic flies results in preferential lethality in male flies. Cytological analysis of mitotic chromosomes reveals that HP1 depletion causes sex-biased chromosomal defects, including telomere fusions. The global levels of specific histone modifications, particularly the hallmarks of active chromatin, are preferentially increased in males as well. Expression analysis revealed that approximately twice as many genes are specifically regulated by HP1 in males compared to females. Furthermore, HP1-regulated genes showed greater enrichment for HP1 binding in males. Taken together, these results reveal that HP1 modulates chromosomal integrity, histone modifications, and transcription in a sex-specific manner.
Keywords: sex-specific, HP1, gender comparison

Overall design

The extraction of total RNA was performed following a standard protocol (www.erin.utoronto.ca/~w3flyma/protocol.htm). Total RNA was isolated from two independent populations of male and female 3rd-instar larva of HP1-21/act-Gal4 and, as controls, larval progenies from line HP1-21 with the genotype y w; +/+; HP1-21/+, and larva with the genotype y w; + /+; +/act-Gal4. In brief, larvaa frozen in liquid nitrogen were homogenized and then resuspended in Trizol reagent by pipetting. The precipitated RNA was washed, and then dissolved in RNase-free water. Five micrograms of total RNA from each experimental sample were reverse-transcribed using the SuperScript Choice cDNA synthesis kit from Stratagene. One microgram of double-stranded cDNA was in vitro-transcribed using the Affymetrix IVT kit and labeled by the incorporation of biotinylated-UTP. Fifteen micrograms of cRNA were then fragmented and hybridized to Affymetrix DGv2 GeneChips as per the manufacturer’s instructions (Affymetrix, Santa Clara CA, USA).

Contributor(s)

Liu L, Ni J, Oakeley EJ, Sun F

Citation(s)

16258543

Submission date

Aug 04, 2005

Last update date

May 04, 2018

Contact name

Edward Oakeley

Organization name

FMI