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Title: RNA from a simple-tandem repeat is required for sperm maturation and male fertility in Drosophila melanogaster

Abstract

Tandemly-repeated DNAs, or satellites, are enriched in heterochromatic regions of eukaryotic genomes and contribute to nuclear structure and function. Some satellites are transcribed, but we lack direct evidence that specific satellite RNAs are required for normal organismal functions. Here, we show satellite RNAs derived from AAGAG tandem repeats are transcribed in many cells throughout Drosophila melanogaster development, enriched in neurons and testes, often localized within heterochromatic regions, and important for viability. Strikingly, we find AAGAG transcripts are necessary for male fertility, and that AAGAG RNA depletion results in defective histone-protamine exchange, sperm maturation and chromatin organization. Since these events happen late in spermatogenesis when the transcripts are not detected, we speculate that AAGAG RNA in primary spermatocytes ‘primes’ post-meiosis steps for sperm maturation. In addition to demonstrating essential functions for AAGAG RNAs, comparisons between closely related Drosophila species suggest that satellites and their transcription evolve quickly to generate new functions.

Authors:
ORCiD logo [1]; ORCiD logo [2]; ORCiD logo [3]; ORCiD logo [3]; ORCiD logo [4]
  1. Univ. of California, Berkeley, CA (United States); Lawrence Berkeley National Lab. (LBNL), Berkeley, CA (United States)
  2. Univ. of California, Berkeley, CA (United States); Lawrence Berkeley National Lab. (LBNL), Berkeley, CA (United States); Univ. of California, Irvine, CA (United States)
  3. National Univ. of Ireland, Galway (Ireland)
  4. Univ. of California, Berkeley, CA (United States);
Publication Date:
Research Org.:
Lawrence Berkeley National Lab. (LBNL), Berkeley, CA (United States)
Sponsoring Org.:
USDOE Office of Science (SC); National Institutes of Health (NIH); National Science Foundation (NSF); Science Foundation Ireland–Health Research Board–Wellcome Trust (SFI-HRB-WT); Science Foundation Ireland –President of Ireland Young Research Award (SFI-PIYRA)
OSTI Identifier:
1591826
Grant/Contract Number:  
[AC02-05CH11231; RO1 GM117420; 00105/Z/12/Z; K99 GM121868; 13/YI/2187]
Resource Type:
Accepted Manuscript
Journal Name:
eLife
Additional Journal Information:
[ Journal Volume: 8]; Journal ID: ISSN 2050-084X
Publisher:
eLife Sciences Publications, Ltd.
Country of Publication:
United States
Language:
English
Subject:
59 BASIC BIOLOGICAL SCIENCES; cell biology; chromosomes and gene expression; satellite RNA; heterochromatin; non-coding RNA; spermatogenesis; histone-protamine exchange; repetitive RNA

Citation Formats

Mills, Wilbur Kyle, Lee, Yuh Chwen G., Kochendoerfer, Antje M., Dunleavy, Elaine M., and Karpen, Gary H. RNA from a simple-tandem repeat is required for sperm maturation and male fertility in Drosophila melanogaster. United States: N. p., 2019. Web. doi:10.7554/eLife.48940.
Mills, Wilbur Kyle, Lee, Yuh Chwen G., Kochendoerfer, Antje M., Dunleavy, Elaine M., & Karpen, Gary H. RNA from a simple-tandem repeat is required for sperm maturation and male fertility in Drosophila melanogaster. United States. doi:10.7554/eLife.48940.
Mills, Wilbur Kyle, Lee, Yuh Chwen G., Kochendoerfer, Antje M., Dunleavy, Elaine M., and Karpen, Gary H. Tue . "RNA from a simple-tandem repeat is required for sperm maturation and male fertility in Drosophila melanogaster". United States. doi:10.7554/eLife.48940. https://www.osti.gov/servlets/purl/1591826.
@article{osti_1591826,
title = {RNA from a simple-tandem repeat is required for sperm maturation and male fertility in Drosophila melanogaster},
author = {Mills, Wilbur Kyle and Lee, Yuh Chwen G. and Kochendoerfer, Antje M. and Dunleavy, Elaine M. and Karpen, Gary H.},
abstractNote = {Tandemly-repeated DNAs, or satellites, are enriched in heterochromatic regions of eukaryotic genomes and contribute to nuclear structure and function. Some satellites are transcribed, but we lack direct evidence that specific satellite RNAs are required for normal organismal functions. Here, we show satellite RNAs derived from AAGAG tandem repeats are transcribed in many cells throughout Drosophila melanogaster development, enriched in neurons and testes, often localized within heterochromatic regions, and important for viability. Strikingly, we find AAGAG transcripts are necessary for male fertility, and that AAGAG RNA depletion results in defective histone-protamine exchange, sperm maturation and chromatin organization. Since these events happen late in spermatogenesis when the transcripts are not detected, we speculate that AAGAG RNA in primary spermatocytes ‘primes’ post-meiosis steps for sperm maturation. In addition to demonstrating essential functions for AAGAG RNAs, comparisons between closely related Drosophila species suggest that satellites and their transcription evolve quickly to generate new functions.},
doi = {10.7554/eLife.48940},
journal = {eLife},
number = ,
volume = [8],
place = {United States},
year = {2019},
month = {11}
}

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Works referenced in this record:

Excavating an Active Site:  The Nucleobase Specificity of Ribonuclease A
journal, November 2000

  • Kelemen, Bradley R.; Schultz, L. Wayne; Sweeney, Rozamond Y.
  • Biochemistry, Vol. 39, Issue 47
  • DOI: 10.1021/bi001862f

High Resolution Fluorescent In Situ Hybridization in Drosophila Embryos and Tissues Using Tyramide Signal Amplification
journal, January 2017

  • Jandura, Allison; Hu, Jack; Wilk, Ronit
  • Journal of Visualized Experiments, Issue 128
  • DOI: 10.3791/56281

AAGAG repeat RNA is an essential component of nuclear matrix in Drosophila
journal, April 2013

  • Pathak, Rashmi; Mamillapalli, Anitha; Rangaraj, Nandini
  • RNA Biology, Vol. 10, Issue 4
  • DOI: 10.4161/rna.24326

Human Centromeres Produce Chromosome-Specific and Array-Specific Alpha Satellite Transcripts that Are Complexed with CENP-A and CENP-C
journal, August 2017


Reactivity and Specificity of RNase T 1 , RNase A, and RNase H toward Oligonucleotides of RNA Containing 8-Oxo-7,8-dihydroguanosine
journal, April 2018


Chromatin dynamics during spermiogenesis
journal, March 2014

  • Rathke, Christina; Baarends, Willy M.; Awe, Stephan
  • Biochimica et Biophysica Acta (BBA) - Gene Regulatory Mechanisms, Vol. 1839, Issue 3
  • DOI: 10.1016/j.bbagrm.2013.08.004

Multiplicity of satellite DNA sequences in Drosophila melanogaster.
journal, February 1986

  • Lohe, A. R.; Brutlag, D. L.
  • Proceedings of the National Academy of Sciences, Vol. 83, Issue 3
  • DOI: 10.1073/pnas.83.3.696

Whole Mount RNA Fluorescent in situ Hybridization of Drosophila Embryos
journal, January 2013

  • Legendre, Félix; Cody, Neal; Iampietro, Carole
  • Journal of Visualized Experiments, Issue 71
  • DOI: 10.3791/50057

Separating from the pack: Molecular mechanisms of Drosophila spermatid individualization
journal, May 2015


Sequence Finishing and Mapping of Drosophila melanogaster Heterochromatin
journal, June 2007


Meiosis in male Drosophila
journal, July 2012

  • McKee, Bruce D.; Yan, Rihui; Tsai, Jui-He
  • Spermatogenesis, Vol. 2, Issue 3
  • DOI: 10.4161/spmg.21800

The modular mechanism of chromocenter formation in Drosophila
journal, February 2019

  • Jagannathan, Madhav; Cummings, Ryan; Yamashita, Yukiko M.
  • eLife, Vol. 8
  • DOI: 10.7554/eLife.43938

The Selfish Segregation Distorter Gene Complex of Drosophila melanogaster
journal, September 2012


Sequence Analysis of a Functional Drosophila Centromere
journal, February 2003


A Short Tandem Repeat-Enriched RNA Assembles a Nuclear Compartment to Control Alternative Splicing and Promote Cell Survival
journal, November 2018


Investigating spermatogenesis in Drosophila melanogaster
journal, June 2014


Preparation of Drosophila Polytene Chromosome Squashes for Antibody Labeling
journal, January 2010

  • Cai, Weili; Jin, Ye; Girton, Jack
  • Journal of Visualized Experiments, Issue 36
  • DOI: 10.3791/1748

Ten principles of heterochromatin formation and function
journal, December 2017

  • Allshire, Robin C.; Madhani, Hiten D.
  • Nature Reviews Molecular Cell Biology, Vol. 19, Issue 4
  • DOI: 10.1038/nrm.2017.119

Direct Evidence of a Role for Heterochromatin in Meiotic Chromosome Segregation
journal, July 1996


Diversity and dynamics of the Drosophila transcriptome
journal, March 2014

  • Brown, James B.; Boley, Nathan; Eisman, Robert
  • Nature, Vol. 512, Issue 7515
  • DOI: 10.1038/nature12962

Prtl99C Acts Together with Protamines and Safeguards Male Fertility in Drosophila
journal, December 2015


Species-Specific Heterochromatin Prevents Mitotic Chromosome Segregation to Cause Hybrid Lethality in Drosophila
journal, October 2009


Multimerization of Drosophila sperm protein Mst77F causes a unique condensed chromatin structure
journal, March 2015

  • Kost, Nils; Kaiser, Sophie; Ostwal, Yogesh
  • Nucleic Acids Research, Vol. 43, Issue 6
  • DOI: 10.1093/nar/gkv015

The Release 6 reference sequence of the Drosophila melanogaster genome
journal, January 2015

  • Hoskins, Roger A.; Carlson, Joseph W.; Wan, Kenneth H.
  • Genome Research, Vol. 25, Issue 3
  • DOI: 10.1101/gr.185579.114

Major satellite repeat RNA stabilize heterochromatin retention of Suv39h enzymes by RNA-nucleosome association and RNA:DNA hybrid formation
journal, August 2017

  • Velazquez Camacho, Oscar; Galan, Carmen; Swist-Rosowska, Kalina
  • eLife, Vol. 6
  • DOI: 10.7554/eLife.25293

New genes as drivers of phenotypic evolution
journal, August 2013

  • Chen, Sidi; Krinsky, Benjamin H.; Long, Manyuan
  • Nature Reviews Genetics, Vol. 14, Issue 9
  • DOI: 10.1038/nrg3521

RNA-dependent stabilization of SUV39H1 at constitutive heterochromatin
journal, August 2017

  • Johnson, Whitney L.; Yewdell, William T.; Bell, Jason C.
  • eLife, Vol. 6
  • DOI: 10.7554/eLife.25299

BRCA1 tumour suppression occurs via heterochromatin-mediated silencing
journal, September 2011

  • Zhu, Quan; Pao, Gerald M.; Huynh, Alexis M.
  • Nature, Vol. 477, Issue 7363
  • DOI: 10.1038/nature10371

A genome-scale shRNA resource for transgenic RNAi in Drosophila
journal, April 2011

  • Ni, Jian-Quan; Zhou, Rui; Czech, Benjamin
  • Nature Methods, Vol. 8, Issue 5
  • DOI: 10.1038/nmeth.1592

Phase separation drives heterochromatin domain formation
journal, June 2017

  • Strom, Amy R.; Emelyanov, Alexander V.; Mir, Mustafa
  • Nature, Vol. 547, Issue 7662
  • DOI: 10.1038/nature22989

Molecular mechanisms of gene regulation during Drosophila spermatogenesis
journal, January 2010


TALE-light imaging reveals maternally guided, H3K9me2/3-independent emergence of functional heterochromatin in Drosophila embryos
journal, February 2016


Tissue, cell type and stage-specific ectopic gene expression and RNAi induction in the Drosophila testis
journal, January 2012


Repetitive centromeric satellite RNA is essential for kinetochore formation and cell division
journal, November 2014

  • Rošić, Silvana; Köhler, Florian; Erhardt, Sylvia
  • The Journal of Cell Biology, Vol. 207, Issue 3
  • DOI: 10.1083/jcb.201404097

Distinct functions of Mst77F and protamines in nuclear shaping and chromatin condensation during Drosophila spermiogenesis
journal, April 2010

  • Rathke, Christina; Barckmann, Bridlin; Burkhard, Silja
  • European Journal of Cell Biology, Vol. 89, Issue 4
  • DOI: 10.1016/j.ejcb.2009.09.001

Ribonuclease III mechanisms of double-stranded RNA cleavage: Ribonuclease III mechanisms of double-stranded RNA cleavage
journal, September 2013

  • Nicholson, Allen W.
  • Wiley Interdisciplinary Reviews: RNA, Vol. 5, Issue 1
  • DOI: 10.1002/wrna.1195

Impact of nucleic acid and methylated H3K9 binding activities of Suv39h1 on its heterochromatin assembly
journal, August 2017


Variable Rates of Simple Satellite Gains across the Drosophila Phylogeny
journal, January 2018

  • Wei, Kevin H. -C; Lower, Sarah E.; Caldas, Ian V.
  • Molecular Biology and Evolution, Vol. 35, Issue 4
  • DOI: 10.1093/molbev/msy005

RNA phase transitions in repeat expansion disorders
journal, May 2017


Origins, evolution, and phenotypic impact of new genes
journal, July 2010


Genome Regulation by Long Noncoding RNAs
journal, July 2012


Mapping simple repeated DNA sequences in heterochromatin of Drosophila melanogaster
journal, November 1993