Fig 1: PWP1 controls the epigenetic status of 5S rDNA. (A) ChIP revealed reduced relative levels of H4K12Ac in the 5S rDNA region following PWP1 depletion in U2OS cells. n=3. Error bars display standard deviation. *P<0.05 (Student's t-test). (B) ChIP revealed increased relative levels of H3K9me2 on 5S rDNA following PWP1 depletion in U2OS cells. n=3. Error bars represent standard deviation. * P<0.05 (Student's t-test). (C) Quantitative RT-PCR analysis of 5S rRNA expression in U2OS cells following PWP1 depletion by siRNA. Human GAPDH was used as a reference gene. n=3. Error bars represent standard deviation. * P<0.05 (Student's t-test).
Fig 2: In Drosophila polytene chromosomes PWP1 binds to the region 56E1-2 that contains the 5S rDNA cluster. (A) The upper panel shows division 57 of chromosome 2R of the Bridges map (Bridges and Bridges, 1939). The lower figure reveals the subdivisions 57C–F in a w1118 polytene chromosome. The red arrow indicates the localization of 5S locus and different tRNAs at the polytene band 56E1–2. (B) Representative images of the subdivisions 56C–F. Immunofluorescence staining with PWP1 antibody and Hoechst revealed the localization of PWP1 in the polytene band 56E1–2. Scale bar: 1 µm. (C) Scheme of the tRNA genes and tandemly repeated 5S rDNA genes as they are represented in Flybase genome browser (http://flybase.org/). The specific tRNA and 5S rDNA clusters are represented in the green and blue boxes, respectively. The blue arrow refers to the neighboring gene OR56a.
Fig 3: Enrichment of PWP1 binding on 5S ribosomal RNA was revealed by chromatin immunoprecipitation (ChIP) in HeLa cells. n=3. Error bars represent standard deviation. *P<0.05 (Student's t-test).
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