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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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Localisation of PtCAF-1 during sexual reproduction by GFP-fusion. PtCAF-1 tagged N-terminally with GFP was expressed at the onset of meiosis and localized in the parental MAC where it remained during skein formation and fragmentation. It later localized in the developing MACs and was present in the new MAC after karyonidal division. (A) Schematic drawing of the life cycle stages of Paramecium tetraurelia. Blue: parental macronucleus; orange: micronuclei, marked with (d) when determined to develop into new MAC. (B) GFP signal. (C) DAPI signal. (D) Merged signals; scale bar: 10 μm.
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Figure 3: Localisation of PtCAF-1 during sexual reproduction by GFP-fusion. PtCAF-1 tagged N-terminally with GFP was expressed at the onset of meiosis and localized in the parental MAC where it remained during skein formation and fragmentation. It later localized in the developing MACs and was present in the new MAC after karyonidal division. (A) Schematic drawing of the life cycle stages of Paramecium tetraurelia. Blue: parental macronucleus; orange: micronuclei, marked with (d) when determined to develop into new MAC. (B) GFP signal. (C) DAPI signal. (D) Merged signals; scale bar: 10 μm.

Mentions: To determine the localization of the PtCAF-1 protein during development, P. tetraurelia was transformed with an N-terminal GFP tagged PTCAF-1 set under the control of a sequence region containing the putative endogenous PTCAF-1 promoter. The localization of PtCAF-1 GFP fusion protein was observed at distinct stages during autogamy by fluorescence microscopy (Figure 3). No GFP signal was detectable during vegetative growth. Upon meiosis the GFP fusion protein was expressed and localized in the parental MAC where it remained during skein formation and in the separated fragments. Later in development the fusion protein accumulated in the new MACs where it persisted throughout MAC development. It was still detectable in the new MACs after karyonidal division. The PtCAF-1 GFP signal decreased with the maturation of the developing MACs. This localization pattern strongly resembles that of Nowa1 (27) reinforcing the hypothesis of PtCAF-1 being a factor in genome rearrangements of P. tetraurelia.


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)

Localisation of PtCAF-1 during sexual reproduction by GFP-fusion. PtCAF-1 tagged N-terminally with GFP was expressed at the onset of meiosis and localized in the parental MAC where it remained during skein formation and fragmentation. It later localized in the developing MACs and was present in the new MAC after karyonidal division. (A) Schematic drawing of the life cycle stages of Paramecium tetraurelia. Blue: parental macronucleus; orange: micronuclei, marked with (d) when determined to develop into new MAC. (B) GFP signal. (C) DAPI signal. (D) Merged signals; scale bar: 10 μm.
© Copyright Policy - creative-commons
Related In: Results  -  Collection

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

Figure 3: Localisation of PtCAF-1 during sexual reproduction by GFP-fusion. PtCAF-1 tagged N-terminally with GFP was expressed at the onset of meiosis and localized in the parental MAC where it remained during skein formation and fragmentation. It later localized in the developing MACs and was present in the new MAC after karyonidal division. (A) Schematic drawing of the life cycle stages of Paramecium tetraurelia. Blue: parental macronucleus; orange: micronuclei, marked with (d) when determined to develop into new MAC. (B) GFP signal. (C) DAPI signal. (D) Merged signals; scale bar: 10 μm.
Mentions: To determine the localization of the PtCAF-1 protein during development, P. tetraurelia was transformed with an N-terminal GFP tagged PTCAF-1 set under the control of a sequence region containing the putative endogenous PTCAF-1 promoter. The localization of PtCAF-1 GFP fusion protein was observed at distinct stages during autogamy by fluorescence microscopy (Figure 3). No GFP signal was detectable during vegetative growth. Upon meiosis the GFP fusion protein was expressed and localized in the parental MAC where it remained during skein formation and in the separated fragments. Later in development the fusion protein accumulated in the new MACs where it persisted throughout MAC development. It was still detectable in the new MACs after karyonidal division. The PtCAF-1 GFP signal decreased with the maturation of the developing MACs. This localization pattern strongly resembles that of Nowa1 (27) reinforcing the hypothesis of PtCAF-1 being a factor in genome rearrangements of P. tetraurelia.

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