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Paramecium tetraurelia chromatin assembly factor-1-like protein PtCAF-1 is involved in RNA-mediated control of DNA elimination.

Ignarski M, Singh A, Swart EC, Arambasic M, Sandoval PY, Nowacki M - Nucleic Acids Res. (2014)

Bottom Line: Gene silencing shows that PtCAF-1 is required for the elimination of transposable elements and a subset of IESs.PTCAF-1 depletion also impairs the selection of germline-specific scnRNAs during development.We identify specific histone modifications appearing during Paramecium development which are strongly reduced in PTCAF-1 depleted cells.

View Article: PubMed Central - PubMed

Affiliation: Institute of Cell Biology, University of Bern, Baltzerstrasse 4, 3012 Bern, Switzerland Graduate School for Cellular and Biomedical Sciences, University of Bern, Freiestrasse 1, 3012 Bern, Switzerland.

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H3K27me3 in control, EZL1 KD, PTIWI01/09 KD and DCL2/3 KD cells. (A) H3K27me3 signal in the parental MAC. (B) H3K27me3 signal in the parental MAC after fragmentation. (C) H3K27me3 signal in the developing MACs. Green: H3K27me3; blue: DAPI; m: parental MAC; dotted circles: developing MAC; scale bar: 10 μm. Control: cells fed with Escherichia coli expressing empty L4440 vector.
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Figure 7: H3K27me3 in control, EZL1 KD, PTIWI01/09 KD and DCL2/3 KD cells. (A) H3K27me3 signal in the parental MAC. (B) H3K27me3 signal in the parental MAC after fragmentation. (C) H3K27me3 signal in the developing MACs. Green: H3K27me3; blue: DAPI; m: parental MAC; dotted circles: developing MAC; scale bar: 10 μm. Control: cells fed with Escherichia coli expressing empty L4440 vector.

Mentions: To verify the specificity of the H3K27me3 antibody we used EZL1 KD cells. Ezl1 is the homologue of Drosophila E(z) and was described in Tetrahymena as developmentally-specific and responsible for H3K27me3 necessary for DNA elimination (38). After EZL1 silencing in Paramecium, H3K27me3 is strongly reduced in the fragments of the parental MAC and is absent from developing MACs (Figure 7). Furthermore, investigation of both PTIWI01/09 and DCL2/3 double knockdown cells revealed the absence of H3K27me3 from the fragments of the parental MAC and from developing MACs (Figure 7).


Paramecium tetraurelia chromatin assembly factor-1-like protein PtCAF-1 is involved in RNA-mediated control of DNA elimination.

Ignarski M, Singh A, Swart EC, Arambasic M, Sandoval PY, Nowacki M - Nucleic Acids Res. (2014)

H3K27me3 in control, EZL1 KD, PTIWI01/09 KD and DCL2/3 KD cells. (A) H3K27me3 signal in the parental MAC. (B) H3K27me3 signal in the parental MAC after fragmentation. (C) H3K27me3 signal in the developing MACs. Green: H3K27me3; blue: DAPI; m: parental MAC; dotted circles: developing MAC; scale bar: 10 μm. Control: cells fed with Escherichia coli expressing empty L4440 vector.
© Copyright Policy - creative-commons
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC4231744&req=5

Figure 7: H3K27me3 in control, EZL1 KD, PTIWI01/09 KD and DCL2/3 KD cells. (A) H3K27me3 signal in the parental MAC. (B) H3K27me3 signal in the parental MAC after fragmentation. (C) H3K27me3 signal in the developing MACs. Green: H3K27me3; blue: DAPI; m: parental MAC; dotted circles: developing MAC; scale bar: 10 μm. Control: cells fed with Escherichia coli expressing empty L4440 vector.
Mentions: To verify the specificity of the H3K27me3 antibody we used EZL1 KD cells. Ezl1 is the homologue of Drosophila E(z) and was described in Tetrahymena as developmentally-specific and responsible for H3K27me3 necessary for DNA elimination (38). After EZL1 silencing in Paramecium, H3K27me3 is strongly reduced in the fragments of the parental MAC and is absent from developing MACs (Figure 7). Furthermore, investigation of both PTIWI01/09 and DCL2/3 double knockdown cells revealed the absence of H3K27me3 from the fragments of the parental MAC and from developing MACs (Figure 7).

Bottom Line: Gene silencing shows that PtCAF-1 is required for the elimination of transposable elements and a subset of IESs.PTCAF-1 depletion also impairs the selection of germline-specific scnRNAs during development.We identify specific histone modifications appearing during Paramecium development which are strongly reduced in PTCAF-1 depleted cells.

View Article: PubMed Central - PubMed

Affiliation: Institute of Cell Biology, University of Bern, Baltzerstrasse 4, 3012 Bern, Switzerland Graduate School for Cellular and Biomedical Sciences, University of Bern, Freiestrasse 1, 3012 Bern, Switzerland.

Show MeSH
Related in: MedlinePlus