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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

The RNase P and RNase MRP catalytic RNAs are not recruited to the activated transgene array. (A) Strand-specific RNA FISH probes were used to visualize the RNase P (RPPH1; a–d) and RNase MRP (RMRP; e–h) catalytic RNAs at the activated transgene array in U2OS 2-6-3 stably expressing YFP-MS2 and rtTA. YFP-MS2 was used to mark the active site (arrows). Yellow lines in enlarged merge insets show the path through which the red and green intensities were measured in the intensity profiles (d, h). Asterisks mark the start of the measured line. Scale bar, 5 μm, 1 μm (enlarged inset). (B) Localization of the RNase P (RPPH1; a–d) and RNase MRP (RMRP; e–h) catalytic RNAs at the activated transgene array in U2OS 2-6-3 cells expressing YFP-Rpp29. Transcription was induced with the activator rtTA (+) Dox. (C) Localization of the RNase P (RPPH1; a–d) and RNase MRP (RMRP; e–h) catalytic RNAs at the activated transgene array in U2OS 2-6-3 cells expressing H3.3-YFP.
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Figure 5: The RNase P and RNase MRP catalytic RNAs are not recruited to the activated transgene array. (A) Strand-specific RNA FISH probes were used to visualize the RNase P (RPPH1; a–d) and RNase MRP (RMRP; e–h) catalytic RNAs at the activated transgene array in U2OS 2-6-3 stably expressing YFP-MS2 and rtTA. YFP-MS2 was used to mark the active site (arrows). Yellow lines in enlarged merge insets show the path through which the red and green intensities were measured in the intensity profiles (d, h). Asterisks mark the start of the measured line. Scale bar, 5 μm, 1 μm (enlarged inset). (B) Localization of the RNase P (RPPH1; a–d) and RNase MRP (RMRP; e–h) catalytic RNAs at the activated transgene array in U2OS 2-6-3 cells expressing YFP-Rpp29. Transcription was induced with the activator rtTA (+) Dox. (C) Localization of the RNase P (RPPH1; a–d) and RNase MRP (RMRP; e–h) catalytic RNAs at the activated transgene array in U2OS 2-6-3 cells expressing H3.3-YFP.

Mentions: To investigate whether the RNase P and/or MRP catalytic RNAs are recruited to the activated array, we used RNA fluorescence in situ hybridization (RNA FISH) to evaluate their localization. It was previously shown that RPPH1, the human RNase P catalytic RNA, localizes in a diffuse pattern throughout the nucleus and that the RNase MRP RNA, RMRP, is enriched in nucleoli (Jacobson et al., 1997). Although we detected RPPH1 and RMRP in their previously reported patterns, neither accumulated at the activated array, marked by YFP-MS2 (Figure 5A). To determine whether we could detect accumulation when either H3.3 or Rpp29, which interacts directly with RPPH1 and RMRP (Marvin and Engelke, 2009), is enriched at the site, we performed RNA FISH in YFP-Rpp29– and H3.3-YFP-expressing cells. The fact that neither RPPH1 nor RMRP accumulated under these conditions (Figure 5, B and C) suggests either that they do not function at the array or that their functions are dysregulated in U2OS cells.


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)

The RNase P and RNase MRP catalytic RNAs are not recruited to the activated transgene array. (A) Strand-specific RNA FISH probes were used to visualize the RNase P (RPPH1; a–d) and RNase MRP (RMRP; e–h) catalytic RNAs at the activated transgene array in U2OS 2-6-3 stably expressing YFP-MS2 and rtTA. YFP-MS2 was used to mark the active site (arrows). Yellow lines in enlarged merge insets show the path through which the red and green intensities were measured in the intensity profiles (d, h). Asterisks mark the start of the measured line. Scale bar, 5 μm, 1 μm (enlarged inset). (B) Localization of the RNase P (RPPH1; a–d) and RNase MRP (RMRP; e–h) catalytic RNAs at the activated transgene array in U2OS 2-6-3 cells expressing YFP-Rpp29. Transcription was induced with the activator rtTA (+) Dox. (C) Localization of the RNase P (RPPH1; a–d) and RNase MRP (RMRP; e–h) catalytic RNAs at the activated transgene array in U2OS 2-6-3 cells expressing H3.3-YFP.
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Related In: Results  -  Collection

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Figure 5: The RNase P and RNase MRP catalytic RNAs are not recruited to the activated transgene array. (A) Strand-specific RNA FISH probes were used to visualize the RNase P (RPPH1; a–d) and RNase MRP (RMRP; e–h) catalytic RNAs at the activated transgene array in U2OS 2-6-3 stably expressing YFP-MS2 and rtTA. YFP-MS2 was used to mark the active site (arrows). Yellow lines in enlarged merge insets show the path through which the red and green intensities were measured in the intensity profiles (d, h). Asterisks mark the start of the measured line. Scale bar, 5 μm, 1 μm (enlarged inset). (B) Localization of the RNase P (RPPH1; a–d) and RNase MRP (RMRP; e–h) catalytic RNAs at the activated transgene array in U2OS 2-6-3 cells expressing YFP-Rpp29. Transcription was induced with the activator rtTA (+) Dox. (C) Localization of the RNase P (RPPH1; a–d) and RNase MRP (RMRP; e–h) catalytic RNAs at the activated transgene array in U2OS 2-6-3 cells expressing H3.3-YFP.
Mentions: To investigate whether the RNase P and/or MRP catalytic RNAs are recruited to the activated array, we used RNA fluorescence in situ hybridization (RNA FISH) to evaluate their localization. It was previously shown that RPPH1, the human RNase P catalytic RNA, localizes in a diffuse pattern throughout the nucleus and that the RNase MRP RNA, RMRP, is enriched in nucleoli (Jacobson et al., 1997). Although we detected RPPH1 and RMRP in their previously reported patterns, neither accumulated at the activated array, marked by YFP-MS2 (Figure 5A). To determine whether we could detect accumulation when either H3.3 or Rpp29, which interacts directly with RPPH1 and RMRP (Marvin and Engelke, 2009), is enriched at the site, we performed RNA FISH in YFP-Rpp29– and H3.3-YFP-expressing cells. The fact that neither RPPH1 nor RMRP accumulated under these conditions (Figure 5, B and C) suggests either that they do not function at the array or that their functions are dysregulated in U2OS cells.

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