75 research outputs found
Postembryonic establishment of megabase-scale gene silencing in nucleolar dominance
Nucleolar dominance is an epigenetic phenomenon in plant and animal genetic hybrids that describes the expression of 45S
ribosomal RNA genes (rRNA genes) inherited from only one progenitor due to the silencing of the other progenitor’s rRNA genes.
rRNA genes are tandemly arrayed at nucleolus organizer regions (NORs) that span millions of basepairs, thus gene silencing in
nucleolar dominance occurs on a scale second only to X-chromosome inactivation in female mammals. In Arabidopsis suecica, the
allotetraploid hybrid of A. thaliana and A. arenosa, theA. thaliana –derived rRNA genes are subjected to nucleolar dominance and
are silenced via repressive chromatin modifications. However, the developmental stage at which nucleolar dominance is
established in A. suecica is currently unknown. We show that nucleolar dominance is not apparent in seedling cotyledons formed
during embryogenesis but becomes progressively established during early postembryonic development in tissues derived from
both the shoot and root apical meristems. The progressive silencing of A. thaliana rRNA genes correlates with the transition of A.
thaliana NORs from a decondensed euchromatic state associated with histone H3 that is trimethylated on lysine 4 (H3K4me3) to
a highly condensed heterochromatic state in which the NORs are associated with H3K9me2 and 5-methylcytosine-enriched
chromocenters. In RNAi-lines in which the histone deacetylases HDA6 and HDT1 are knocked down, the developmentally
regulated condensation and inactivation of A. thaliana NORs is disrupted. Collectively, these data demonstrate that HDA6 and
HDT1 function in the postembryonic establishment of nucleolar dominance, a process which recurs in each generatio
Autoregulation of the Drosophila Noncoding roX1 RNA Gene
Most genes along the male single X chromosome in Drosophila are hypertranscribed about two-fold relative to each of the two female X chromosomes. This is accomplished by the MSL (male-specific lethal) complex that acetylates histone H4 at lysine 16. The MSL complex contains two large noncoding RNAs, roX1 (RNA on X) and roX2, that help target chromatin modifying enzymes to the X. The roX RNAs are functionally redundant but differ in size, sequence, and transcriptional control. We wanted to find out how roX1 production is regulated. Ectopic DC can be induced in wild-type (roX1+ roX2+) females if we provide a heterologous source of MSL2. However, in the absence of roX2, we found that roX1 expression failed to come on reliably. Using an in situ hybridization probe that is specific only to endogenous roX1, we found that expression was restored if we introduced either roX2 or a truncated but functional version of roX1. This shows that pre-existing roX RNA is required to positively autoregulate roX1 expression. We also observed massive cis spreading of the MSL complex from the site of roX1 transcription at its endogenous location on the X chromosome. We propose that retention of newly assembled MSL complex around the roX gene is needed to drive sustained transcription and that spreading into flanking chromatin contributes to the X chromosome targeting specificity. Finally, we found that the gene encoding the key male-limited protein subunit, msl2, is transcribed predominantly during DNA replication. This suggests that new MSL complex is made as the chromatin template doubles. We offer a model describing how the production of roX1 and msl2, two key components of the MSL complex, are coordinated to meet the dosage compensation demands of the male cell
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