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RNase P protein subunit Rpp29 represses histone H3.3 nucleosome deposition.

Newhart A, Powers SL, Shastrula PK, Sierra I, Joo LM, Hayden JE, Cohen AR, Janicki SM - Mol. Biol. Cell (2016)

Bottom Line: Rpp29 is a protein subunit of RNase P.Of the other subunits, POP1 and Rpp21 are similarly recruited suggesting that a variant of RNase P regulates H3.3 chromatin assembly.Rpp29 knockdown increases H3.3 chromatin incorporation, which suggests that Rpp29 represses H3.3 nucleosome deposition, a finding with implications for epigenetic regulation.

View Article: PubMed Central - PubMed

Affiliation: Molecular and Cellular Oncogenesis Program, Wistar Institute, Philadelphia, PA 19104.

No MeSH data available.


Related in: MedlinePlus

Rpp29 and POP1 repress transcription from the transgene array. (A) qRT-PCR analysis of Rpp29 and POP1 mRNA levels in U2OS 2-6-3 cells stably expressing YFP-MS2 and rtTA cells after shRNA knockdown. SDs are shown in the form of error bars; p values were calculated using an unpaired t test (n = 3). (B) Strand-specific qRT-PCR analysis of total RNA isolated 0 and 3 h after activation in U2OS 263 cells stably expressing YFP-MS2 and rtTA after shRNA knockdown. A primer pair in rabbit β-globin exon 3 was used for PCR. (C) Measurement of the intensity of H3.3-YFP recruited to the activated transgene array in U2OS 2-6-3 cells after control (pLKO.1; n = 30) and Rpp29 (n = 31) knockdown.
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Figure 7: Rpp29 and POP1 repress transcription from the transgene array. (A) qRT-PCR analysis of Rpp29 and POP1 mRNA levels in U2OS 2-6-3 cells stably expressing YFP-MS2 and rtTA cells after shRNA knockdown. SDs are shown in the form of error bars; p values were calculated using an unpaired t test (n = 3). (B) Strand-specific qRT-PCR analysis of total RNA isolated 0 and 3 h after activation in U2OS 263 cells stably expressing YFP-MS2 and rtTA after shRNA knockdown. A primer pair in rabbit β-globin exon 3 was used for PCR. (C) Measurement of the intensity of H3.3-YFP recruited to the activated transgene array in U2OS 2-6-3 cells after control (pLKO.1; n = 30) and Rpp29 (n = 31) knockdown.

Mentions: To test the hypothesis that Rpp29 represses transcription, we knocked down Rpp29, as well as POP1, in the U2OS cell line and measured S and AS RNA levels. Figure 7A shows the depletion of Rpp29 and POP1 mRNA in short hairpin RNA (shRNA)–expressing cells. Knockdown of Rpp29 significantly increased levels of both S (activated cells) and AS RNA (inactive and activated cells), whereas POP1 knockdown increased S RNA levels (activated cells; Figure 7B). This result supports the hypothesis that Rpp29, as well as POP1, represses transcription from the transgene array.


RNase P protein subunit Rpp29 represses histone H3.3 nucleosome deposition.

Newhart A, Powers SL, Shastrula PK, Sierra I, Joo LM, Hayden JE, Cohen AR, Janicki SM - Mol. Biol. Cell (2016)

Rpp29 and POP1 repress transcription from the transgene array. (A) qRT-PCR analysis of Rpp29 and POP1 mRNA levels in U2OS 2-6-3 cells stably expressing YFP-MS2 and rtTA cells after shRNA knockdown. SDs are shown in the form of error bars; p values were calculated using an unpaired t test (n = 3). (B) Strand-specific qRT-PCR analysis of total RNA isolated 0 and 3 h after activation in U2OS 263 cells stably expressing YFP-MS2 and rtTA after shRNA knockdown. A primer pair in rabbit β-globin exon 3 was used for PCR. (C) Measurement of the intensity of H3.3-YFP recruited to the activated transgene array in U2OS 2-6-3 cells after control (pLKO.1; n = 30) and Rpp29 (n = 31) knockdown.
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Related In: Results  -  Collection

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Figure 7: Rpp29 and POP1 repress transcription from the transgene array. (A) qRT-PCR analysis of Rpp29 and POP1 mRNA levels in U2OS 2-6-3 cells stably expressing YFP-MS2 and rtTA cells after shRNA knockdown. SDs are shown in the form of error bars; p values were calculated using an unpaired t test (n = 3). (B) Strand-specific qRT-PCR analysis of total RNA isolated 0 and 3 h after activation in U2OS 263 cells stably expressing YFP-MS2 and rtTA after shRNA knockdown. A primer pair in rabbit β-globin exon 3 was used for PCR. (C) Measurement of the intensity of H3.3-YFP recruited to the activated transgene array in U2OS 2-6-3 cells after control (pLKO.1; n = 30) and Rpp29 (n = 31) knockdown.
Mentions: To test the hypothesis that Rpp29 represses transcription, we knocked down Rpp29, as well as POP1, in the U2OS cell line and measured S and AS RNA levels. Figure 7A shows the depletion of Rpp29 and POP1 mRNA in short hairpin RNA (shRNA)–expressing cells. Knockdown of Rpp29 significantly increased levels of both S (activated cells) and AS RNA (inactive and activated cells), whereas POP1 knockdown increased S RNA levels (activated cells; Figure 7B). This result supports the hypothesis that Rpp29, as well as POP1, represses transcription from the transgene array.

Bottom Line: Rpp29 is a protein subunit of RNase P.Of the other subunits, POP1 and Rpp21 are similarly recruited suggesting that a variant of RNase P regulates H3.3 chromatin assembly.Rpp29 knockdown increases H3.3 chromatin incorporation, which suggests that Rpp29 represses H3.3 nucleosome deposition, a finding with implications for epigenetic regulation.

View Article: PubMed Central - PubMed

Affiliation: Molecular and Cellular Oncogenesis Program, Wistar Institute, Philadelphia, PA 19104.

No MeSH data available.


Related in: MedlinePlus