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Genome-Wide Distribution of Nascent Transcripts in Sperm DNA, Products of a Late Wave of General Transcription

Mature spermatozoa contain a whole repertoire of the various classes of cellular RNAs, both coding and non-coding. It was hypothesized that after fertilization they might impact development, a claim supported by experimental evidence in various systems. Despite the current increasing interest in the...

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Autores principales: Kianmehr, Leila, Khazali, Homayoun, Rajabi-Maham, Hassan, Sharifi-Zarchi, Ali, Cuzin, François, Rassoulzadegan, Minoo
Formato: Online Artículo Texto
Lenguaje:English
Publicado: MDPI 2019
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC6830337/
https://www.ncbi.nlm.nih.gov/pubmed/31623338
http://dx.doi.org/10.3390/cells8101196
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author Kianmehr, Leila
Khazali, Homayoun
Rajabi-Maham, Hassan
Sharifi-Zarchi, Ali
Cuzin, François
Rassoulzadegan, Minoo
author_facet Kianmehr, Leila
Khazali, Homayoun
Rajabi-Maham, Hassan
Sharifi-Zarchi, Ali
Cuzin, François
Rassoulzadegan, Minoo
author_sort Kianmehr, Leila
collection PubMed
description Mature spermatozoa contain a whole repertoire of the various classes of cellular RNAs, both coding and non-coding. It was hypothesized that after fertilization they might impact development, a claim supported by experimental evidence in various systems. Despite the current increasing interest in the transgenerational maintenance of epigenetic traits and their possible determination by RNAs, little remains known about conservation in sperm and across generations and the specificities and mechanisms involved in transgenerational maintenance. We identified two distinct fractions of RNAs in mature mouse sperm, one readily extracted in the aqueous phase of the classical TRIzol procedure and a distinct fraction hybridized with homologous DNA in DNA-RNA complexes recovered from the interface, purified after DNase hydrolysis and analyzed by RNA-seq methodology. This DNA-associated RNA (D RNA) was found to represent as much as half of the cell contents in differentiated sperm, in which a major part of the cytoplasmic material has been discarded. Stable complexes were purified free of proteins and identified as hybrids (R-loops) on the basis of their sensitivity to RNase H hydrolysis. Further analysis by RNA-seq identified transcripts from all the coding and non-coding regions of the genome, thus revealing an extensive wave of transcription, prior to or concomitant with the terminal compaction of the chromatin.
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spelling pubmed-68303372019-11-20 Genome-Wide Distribution of Nascent Transcripts in Sperm DNA, Products of a Late Wave of General Transcription Kianmehr, Leila Khazali, Homayoun Rajabi-Maham, Hassan Sharifi-Zarchi, Ali Cuzin, François Rassoulzadegan, Minoo Cells Article Mature spermatozoa contain a whole repertoire of the various classes of cellular RNAs, both coding and non-coding. It was hypothesized that after fertilization they might impact development, a claim supported by experimental evidence in various systems. Despite the current increasing interest in the transgenerational maintenance of epigenetic traits and their possible determination by RNAs, little remains known about conservation in sperm and across generations and the specificities and mechanisms involved in transgenerational maintenance. We identified two distinct fractions of RNAs in mature mouse sperm, one readily extracted in the aqueous phase of the classical TRIzol procedure and a distinct fraction hybridized with homologous DNA in DNA-RNA complexes recovered from the interface, purified after DNase hydrolysis and analyzed by RNA-seq methodology. This DNA-associated RNA (D RNA) was found to represent as much as half of the cell contents in differentiated sperm, in which a major part of the cytoplasmic material has been discarded. Stable complexes were purified free of proteins and identified as hybrids (R-loops) on the basis of their sensitivity to RNase H hydrolysis. Further analysis by RNA-seq identified transcripts from all the coding and non-coding regions of the genome, thus revealing an extensive wave of transcription, prior to or concomitant with the terminal compaction of the chromatin. MDPI 2019-10-03 /pmc/articles/PMC6830337/ /pubmed/31623338 http://dx.doi.org/10.3390/cells8101196 Text en © 2019 by the authors. Licensee MDPI, Basel, Switzerland. This article is an open access article distributed under the terms and conditions of the Creative Commons Attribution (CC BY) license (http://creativecommons.org/licenses/by/4.0/).
spellingShingle Article
Kianmehr, Leila
Khazali, Homayoun
Rajabi-Maham, Hassan
Sharifi-Zarchi, Ali
Cuzin, François
Rassoulzadegan, Minoo
Genome-Wide Distribution of Nascent Transcripts in Sperm DNA, Products of a Late Wave of General Transcription
title Genome-Wide Distribution of Nascent Transcripts in Sperm DNA, Products of a Late Wave of General Transcription
title_full Genome-Wide Distribution of Nascent Transcripts in Sperm DNA, Products of a Late Wave of General Transcription
title_fullStr Genome-Wide Distribution of Nascent Transcripts in Sperm DNA, Products of a Late Wave of General Transcription
title_full_unstemmed Genome-Wide Distribution of Nascent Transcripts in Sperm DNA, Products of a Late Wave of General Transcription
title_short Genome-Wide Distribution of Nascent Transcripts in Sperm DNA, Products of a Late Wave of General Transcription
title_sort genome-wide distribution of nascent transcripts in sperm dna, products of a late wave of general transcription
topic Article
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC6830337/
https://www.ncbi.nlm.nih.gov/pubmed/31623338
http://dx.doi.org/10.3390/cells8101196
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