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      Dynamic sex chromosome expression in Drosophila male germ cells

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          Abstract

          Given their copy number differences and unique modes of inheritance, the evolved gene content and expression of sex chromosomes is unusual. In many organisms the X and Y chromosomes are inactivated in spermatocytes, possibly as a defense mechanism against insertions into unpaired chromatin. In addition to current sex chromosomes, Drosophila has a small gene-poor X-chromosome relic (4 th) that re-acquired autosomal status. Here we use single cell RNA-Seq on fly larvae to demonstrate that the single X and pair of 4 th chromosomes are specifically inactivated in primary spermatocytes, based on measuring all genes or a set of broadly expressed genes in testis we identified. In contrast, genes on the single Y chromosome become maximally active in primary spermatocytes. Reduced X transcript levels are due to failed activation of RNA-Polymerase-II by phosphorylation of Serine 2 and 5.

          Abstract

          Sex chromosome gene content and expression is unusual. Here the authors use single cell RNA-Seq on Drosophila larvae to demonstrate that the single X and pair of 4th chromosomes are specifically inactivated in primary spermatocytes, while genes on the single Y chromosome become maximally active in primary spermatocytes.

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          The Sequence Alignment/Map format and SAMtools

          Summary: The Sequence Alignment/Map (SAM) format is a generic alignment format for storing read alignments against reference sequences, supporting short and long reads (up to 128 Mbp) produced by different sequencing platforms. It is flexible in style, compact in size, efficient in random access and is the format in which alignments from the 1000 Genomes Project are released. SAMtools implements various utilities for post-processing alignments in the SAM format, such as indexing, variant caller and alignment viewer, and thus provides universal tools for processing read alignments. Availability: http://samtools.sourceforge.net Contact: rd@sanger.ac.uk
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            Cutadapt removes adapter sequences from high-throughput sequencing reads

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              featureCounts: an efficient general purpose program for assigning sequence reads to genomic features.

              Next-generation sequencing technologies generate millions of short sequence reads, which are usually aligned to a reference genome. In many applications, the key information required for downstream analysis is the number of reads mapping to each genomic feature, for example to each exon or each gene. The process of counting reads is called read summarization. Read summarization is required for a great variety of genomic analyses but has so far received relatively little attention in the literature. We present featureCounts, a read summarization program suitable for counting reads generated from either RNA or genomic DNA sequencing experiments. featureCounts implements highly efficient chromosome hashing and feature blocking techniques. It is considerably faster than existing methods (by an order of magnitude for gene-level summarization) and requires far less computer memory. It works with either single or paired-end reads and provides a wide range of options appropriate for different sequencing applications. featureCounts is available under GNU General Public License as part of the Subread (http://subread.sourceforge.net) or Rsubread (http://www.bioconductor.org) software packages.
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                Author and article information

                Contributors
                briano@niddk.nih.gov
                Journal
                Nat Commun
                Nat Commun
                Nature Communications
                Nature Publishing Group UK (London )
                2041-1723
                9 February 2021
                9 February 2021
                2021
                : 12
                : 892
                Affiliations
                [1 ]GRID grid.94365.3d, ISNI 0000 0001 2297 5165, Laboratory of Cellular and Developmental Biology, National Institute of Diabetes and Kidney and Digestive Diseases, , National Institutes of Health, ; Bethesda, MD 20892 USA
                [2 ]GRID grid.21107.35, ISNI 0000 0001 2171 9311, Department of Cell Biology, , Johns Hopkins University School of Medicine, ; 725 N. Wolfe Street, Baltimore, MD 21205 USA
                [3 ]GRID grid.94365.3d, ISNI 0000 0001 2297 5165, Cell Biology and Physiology Center, National Heart, , Lung and Blood Institute, National Institutes of Health, ; Bethesda, MD 20892 USA
                [4 ]GRID grid.11899.38, ISNI 0000 0004 1937 0722, Department of Genetics and Evolutionary Biology, Institute of Biosciences, , University of São Paulo, ; SP 05508-090 São Paulo, Brazil
                [5 ]GRID grid.410543.7, ISNI 0000 0001 2188 478X, Instituto de Biociências/IB, Departamento de Biologia Geral e Aplicada, , UNESP—Universidade Estadual Paulista, Rio Claro, ; São Paulo, 13506-900 Brazil
                [6 ]GRID grid.94365.3d, ISNI 0000 0001 2297 5165, Laboratory of Muscle Stem Cells and Gene Regulation, National Institute of Arthritis and Musculoskeletal and Skin Diseases, , National Institutes of Health, ; Bethesda, MD 20892 USA
                [7 ]GRID grid.94365.3d, ISNI 0000 0001 2297 5165, Genomics Core, National Institute of Diabetes and Kidney and Digestive Diseases, , National Institutes of Health, ; Bethesda, MD 20892 USA
                [8 ]GRID grid.8993.b, ISNI 0000 0004 1936 9457, Present Address: Department of Evolutionary Biology and Department of Organismal Biology, Systematic Biology, Evolutionary Biology Centre, , Uppsala University, ; 75236 Uppsala, Sweden
                [9 ]GRID grid.16753.36, ISNI 0000 0001 2299 3507, Present Address: Department of Pharmacology, Feinberg School of Medicine, , Northwestern University, ; Chicago, IL 60611 USA
                Author information
                http://orcid.org/0000-0002-2732-449X
                http://orcid.org/0000-0002-6417-0637
                http://orcid.org/0000-0002-1306-2820
                http://orcid.org/0000-0002-3517-5973
                http://orcid.org/0000-0002-9284-3675
                http://orcid.org/0000-0002-3455-4891
                Article
                20897
                10.1038/s41467-021-20897-y
                7873209
                33563972
                8917ef77-e603-4914-9e81-6a9b0c80a0e3
                © This is a U.S. government work and not under copyright protection in the U.S.; foreign copyright protection may apply 2021

                Open Access This article is licensed under a Creative Commons Attribution 4.0 International License, which permits use, sharing, adaptation, distribution and reproduction in any medium or format, as long as you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons license, and indicate if changes were made. The images or other third party material in this article are included in the article’s Creative Commons license, unless indicated otherwise in a credit line to the material. If material is not included in the article’s Creative Commons license and your intended use is not permitted by statutory regulation or exceeds the permitted use, you will need to obtain permission directly from the copyright holder. To view a copy of this license, visit http://creativecommons.org/licenses/by/4.0/.

                History
                : 11 April 2020
                : 22 December 2020
                Funding
                Funded by: FundRef https://doi.org/10.13039/100000062, U.S. Department of Health & Human Services | NIH | National Institute of Diabetes and Digestive and Kidney Diseases (National Institute of Diabetes & Digestive & Kidney Diseases);
                Award ID: DK015600
                Award Recipient :
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                © The Author(s) 2021

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                drosophila,rna sequencing,spermatogenesis,evolutionary developmental biology,development

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