A new strategy for isolating genes controlling dosage compensation in Drosophila using a simple epigenetic mosaic eye phenotype.
BACKGROUND
The Drosophila Male Specific Lethal (MSL) complex contains chromatin modifying enzymes and non-coding roX RNA. It paints the male X at hundreds of bands where it acetylates histone H4 at lysine 16. This epigenetic mark increases expression from the single male X chromosome approximately twofold above what gene-specific factors produce from each female X chromosome. This equalises X-linked gene expression between the sexes. Previous screens for components of dosage compensation relied on a distinctive male-specific lethal phenotype.


RESULTS
Here, we report a new strategy relying upon an unusual male-specific mosaic eye pigmentation phenotype produced when the MSL complex acts upon autosomal roX1 transgenes. Screening the second chromosome identified at least five loci, two of which are previously described components of the MSL complex. We focused our analysis on the modifier alleles of MSL1 and MLE (for 'maleless'). The MSL1 lesions are not simple nulls, but rather alter the PEHE domain that recruits the MSL3 chromodomain and MOF ('males absent on first') histone acetyltransferase subunits to the complex. These mutants are compromised in their ability to recruit MSL3 and MOF, dosage compensate the X, and support long distance spreading from roX1 transgenes. Yet, paradoxically, they were isolated because they somehow increase MSL complex activity immediately around roX1 transgenes in combination with wild-type MSL1 subunits.


CONCLUSIONS
We propose that these diverse phenotypes arise from perturbations in assembly of MSL subunits onto nascent roX transcripts. This strategy is a promising alternative route for identifying previously unknown components of the dosage compensation pathway and novel alleles of known MSL proteins.
Program in Developmental Biology, Baylor College of Medicine, Houston, TX, USA.
Mahalakshmi
M
Prabhakaran
Richard L
RL
Kelley
D000818:Animals; D001483:Base Sequence; D015152:Blotting, Northern; D015153:Blotting, Western; D017931:DNA Primers / Q000235:genetics; D004303:Dosage Compensation, Genetic / Q000235:genetics*; D029721:Drosophila Proteins / Q000378:metabolism*; D004331:Drosophila melanogaster / Q000235:genetics*; D005786:Gene Expression Regulation / Q000235:genetics*; D005816:Genetic Complementation Test; D051548:Histone Acetyltransferases / Q000378:metabolism; D047468:Immunoprecipitation; D008297:Male; D008969:Molecular Sequence Data; D016296:Mutagenesis; D009687:Nuclear Proteins / Q000378:metabolism*; D009799:Ocular Physiological Phenomena / Q000235:genetics; D010858:Pigmentation / Q000235:genetics / Q000502:physiology; D017422:Sequence Analysis, DNA; D012737:Sex Factors; D014157:Transcription Factors / Q000378:metabolism*; D014960:X Chromosome / Q000235:genetics* / Q000378:metabolism
D017931:DNA Primers; D029721:Drosophila Proteins; D009687:Nuclear Proteins; D014157:Transcription Factors; C082132:msl-1 protein, Drosophila; C454144:roX1 protein, Drosophila; D051548:Histone Acetyltransferases; C111063:mof protein, Drosophila