Ordered assembly of roX RNAs into MSL complexes on the dosage-compensated X chromosome in Drosophila.
BACKGROUND
In the male Drosophila, the X chromosome is transcriptionally upregulated to achieve dosage compensation, in a process that depends on association of the MSL proteins with the X chromosome. A role for non-coding RNAs has been suggested in recent studies. The roX1 and roX2 RNAs are male-specific, non-coding RNAs that are produced by, and also found associated with, the dosage-compensated male X chromosome. Whether roX RNAs are physically part of the MSL complex has not been resolved.


RESULTS
We found that roX RNAs colocalize with the MSL proteins and are highly unstable unless the MSL complex is coexpressed, suggesting a physical interaction. We were able to immunoprecipitate roX2 RNA from male tissue-culture cells with antibodies to the proteins Msl1 and Mle, consistent with an integral association with MSL complexes. Localization of roX1 and roX2 RNAs in mutants indicated an order of MSL-complex assembly in which roX2 RNA is incorporated early in a process requiring the Mle helicase. We also found that the roX2 gene, like roX1, is a nucleation site for MSL complex spreading into flanking chromatin in cis.


CONCLUSIONS
Our results support a model in which MSL proteins assemble at specific chromatin entry sites (including the roX1 and roX2 genes); the roX RNAs join the complex at their sites of synthesis; and complete complexes spread in cis to dosage compensate most genes on the X chromosome.
Meller
V H
VH
Department of Molecular and Cellular Biology, Department of Biology, Baylor College of Medicine, Tufts University, Houston, Medford 77030, 02155, USA.
Gordadze
P R
PR
Park
Y
Y
Chu
X
X
Stuckenholz
C
C
Kelley
R L
RL
Kuroda
M I
MI
D000818:Animals; D002843:Chromatin / Q000235:genetics / Q000378:metabolism; D004303:Dosage Compensation, Genetic; D004330:Drosophila / Q000235:genetics / Q000378:metabolism; D017344:Genes, Insect; D017404:In Situ Hybridization, Fluorescence; D008297:Male; D009687:Nuclear Proteins / Q000235:genetics / Q000378:metabolism*; D011233:Precipitin Tests; D012313:RNA / Q000235:genetics / Q000378:metabolism*; D019076:Transgenes; D014960:X Chromosome / Q000235:genetics* / Q000378:metabolism
D002843:Chromatin; D009687:Nuclear Proteins; D012313:RNA