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Characterization of MRP RNA–protein interactions within the perinucleolar compartment
The perinucleolar compartment (PNC) forms in cancer cells and is highly enriched with a subset of polymerase III RNAs and RNA-binding proteins. Here we report that PNC components mitochondrial RNA–processing (MRP) RNA, pyrimidine tract–binding protein (PTB), and CUG-binding protein (CUGBP) interact...
Autores principales: | , , , , , , |
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Formato: | Texto |
Lenguaje: | English |
Publicado: |
The American Society for Cell Biology
2011
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Materias: | |
Acceso en línea: | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC3057709/ https://www.ncbi.nlm.nih.gov/pubmed/21233287 http://dx.doi.org/10.1091/mbc.E10-09-0768 |
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author | Pollock, Callie Daily, Kelly Nguyen, Van Trung Wang, Chen Lewandowska, Marzena Anna Bensaude, Olivier Huang, Sui |
author_facet | Pollock, Callie Daily, Kelly Nguyen, Van Trung Wang, Chen Lewandowska, Marzena Anna Bensaude, Olivier Huang, Sui |
author_sort | Pollock, Callie |
collection | PubMed |
description | The perinucleolar compartment (PNC) forms in cancer cells and is highly enriched with a subset of polymerase III RNAs and RNA-binding proteins. Here we report that PNC components mitochondrial RNA–processing (MRP) RNA, pyrimidine tract–binding protein (PTB), and CUG-binding protein (CUGBP) interact in vivo, as demonstrated by coimmunoprecipitation and RNA pull-down experiments. Glycerol gradient analyses show that this complex is large and sediments at a different fraction from known MRP RNA–containing complexes, the MRP ribonucleoprotein ribozyme and human telomerase reverse transcriptase. Tethering PNC components to a LacO locus recruits other PNC components, further confirming the in vivo interactions. These interactions are present both in PNC-containing and -lacking cells. High-resolution localization analyses demonstrate that MRP RNA, CUGBP, and PTB colocalize at the PNC as a reticulated network, intertwining with newly synthesized RNA. Furthermore, green fluorescent protein (GFP)–PTB and GFP-CUGBP show a slower rate of fluorescence recovery after photobleaching at the PNC than in the nucleoplasm, illustrating the different molecular interaction of the complexes associated with the PNC. These findings support a working model in which the MRP RNA–protein complex becomes nucleated at the PNC in cancer cells and may play a role in gene expression regulation at the DNA locus that associates with the PNC. |
format | Text |
id | pubmed-3057709 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2011 |
publisher | The American Society for Cell Biology |
record_format | MEDLINE/PubMed |
spelling | pubmed-30577092011-05-30 Characterization of MRP RNA–protein interactions within the perinucleolar compartment Pollock, Callie Daily, Kelly Nguyen, Van Trung Wang, Chen Lewandowska, Marzena Anna Bensaude, Olivier Huang, Sui Mol Biol Cell Articles The perinucleolar compartment (PNC) forms in cancer cells and is highly enriched with a subset of polymerase III RNAs and RNA-binding proteins. Here we report that PNC components mitochondrial RNA–processing (MRP) RNA, pyrimidine tract–binding protein (PTB), and CUG-binding protein (CUGBP) interact in vivo, as demonstrated by coimmunoprecipitation and RNA pull-down experiments. Glycerol gradient analyses show that this complex is large and sediments at a different fraction from known MRP RNA–containing complexes, the MRP ribonucleoprotein ribozyme and human telomerase reverse transcriptase. Tethering PNC components to a LacO locus recruits other PNC components, further confirming the in vivo interactions. These interactions are present both in PNC-containing and -lacking cells. High-resolution localization analyses demonstrate that MRP RNA, CUGBP, and PTB colocalize at the PNC as a reticulated network, intertwining with newly synthesized RNA. Furthermore, green fluorescent protein (GFP)–PTB and GFP-CUGBP show a slower rate of fluorescence recovery after photobleaching at the PNC than in the nucleoplasm, illustrating the different molecular interaction of the complexes associated with the PNC. These findings support a working model in which the MRP RNA–protein complex becomes nucleated at the PNC in cancer cells and may play a role in gene expression regulation at the DNA locus that associates with the PNC. The American Society for Cell Biology 2011-03-15 /pmc/articles/PMC3057709/ /pubmed/21233287 http://dx.doi.org/10.1091/mbc.E10-09-0768 Text en © 2011 Pollock et al. This article is distributed by The American Society for Cell Biology under license from the author(s). Two months after publication it is available to the public under an Attribution–Noncommercial–Share Alike 3.0 Unported Creative Commons License (http://creativecommons.org/licenses/by-nc-sa/3.0). “ASCB®,“ “The American Society for Cell Biology®,” and “Molecular Biology of the Cell®” are registered trademarks of The American Society of Cell Biology. |
spellingShingle | Articles Pollock, Callie Daily, Kelly Nguyen, Van Trung Wang, Chen Lewandowska, Marzena Anna Bensaude, Olivier Huang, Sui Characterization of MRP RNA–protein interactions within the perinucleolar compartment |
title | Characterization of MRP RNA–protein interactions within the perinucleolar compartment |
title_full | Characterization of MRP RNA–protein interactions within the perinucleolar compartment |
title_fullStr | Characterization of MRP RNA–protein interactions within the perinucleolar compartment |
title_full_unstemmed | Characterization of MRP RNA–protein interactions within the perinucleolar compartment |
title_short | Characterization of MRP RNA–protein interactions within the perinucleolar compartment |
title_sort | characterization of mrp rna–protein interactions within the perinucleolar compartment |
topic | Articles |
url | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC3057709/ https://www.ncbi.nlm.nih.gov/pubmed/21233287 http://dx.doi.org/10.1091/mbc.E10-09-0768 |
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