Crossroads between transposons and gene regulation

09:05-09:40 Sleeping with the enemy: Methylation-seeking retrotransposons across eukaryotic lineages

Dr Alex de Mendoza, ARC CoE Plant Energy Biology, The University of Western Australia, Australia

Abstract

Genomic cytosine methylation is a major mechanism to transcriptionally silence transposable elements across eukaryotes. Therefore, transposable elements are expected to evolve mechanisms to evade methylation to remain active in the host genomes. However, the discovery of several retrotransposons that have hijacked DNA methylation through the acquisition of host genes challenges this expectation. In one case, three distinct retrotransposon classes have acquired cytosine DNA methyltransferases, most spectacularly in the genomes of Symbiodinium dinoflagellates, where hundreds to thousands of copies of these retrotransposons are likely to have influenced the host’s exceptional epigenomic landscape. In another case, a group of arthropod retrotransposons have acquired a functionally active Methyl-CpG Binding Domain. This domain guides the retrotransposon integration into methylated CpG-rich regions, which correspond to repetitive regions of the genome, thus likely decreasing the potentially harmful effects of new insertions. These illustrate the convoluted arms race between transposable elements and their host genomes. Both cases exemplify the delicate balance between expression and silencing that retrotransposons need to achieve in order to co-exist with their host. These serve as the first examples of how transposons can domesticate host genes central to epigenome regulation to their own advantage.

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09:40-10:15 Transposable element control during germline development in Drosophila

Dr Felipe Karam Teixeira, University of Cambridge, UK

10:15-10:45 Tea/Coffee

11:00-12:20 Impact of transposable elements on human gene regulation

Professor Joanna Wysocka, Stanford University, USA

Abstract

Professor Wysocka’s team is interested in understanding how TEs can serve as a substrate for evolution of novel regulatory elements and functions in the primate lineage. During human pre-implantation development many TEs escape silencing and are transcribed at discreet stages, with most stage-specific regulation occurring at the class of LTR retrotransposons called endogenous retroviruses (ERVs), which are remnants of ancient retroviral infections. One such ERV class, called HERVK, retained coding potential and gives rise to the viral-like particles and proteins that are readily detected in human blastocysts and can influence innate immune response. The team discovered that HERVK LTR elements, called LTR5HS, function as ape-specific enhancers and regulate expression of nearly 300 human embryonic genes. A flip side to TEs serving as a playground for regulatory innovation is that mobile transposons are parasitic elements that pose an ongoing threat to the genome, and thus must be controlled by the host cell. The non-LTR retrotransposon LINE-1 (L1) is the only currently mobile autonomous TE in humans. In collaboration with the Bassik lab at Stanford, the team performed first genome-wide screen for regulators of L1 retrotransposition and identified ~150 human genes that either activate or repress L1 retrotransposition in human cells. Through this screen, the researchers discovered a novel mechanism dedicated to transcriptional silencing of evolutionarily young, full-length L1s immersed within transcriptionally permissive euchromatic environment, and showed that this silencing pathway also has a collateral effect on the host gene expression programs. Professor Wysocka will discuss these findings and talk about her team ongoing work.

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11:20-11:55 The impact of transposable element invasions on the evolution of human neuronal gene expression

Dr Frank Jacobs, University of Amsterdam, the Netherlands

Abstract

Approximately half of our genome consists of repetitive DNA sequences, mainly contained in transposable elements (TEs). TEs are retrovirus-derived DNA elements which copy-pasted themselves throughout our genome during vertebrate evolution. TE invasions have been a major driver of genome evolution but it remains elusive to which extent TEs have influenced how genes in the human genome are regulated. In previous work (Jacobs et al, 2014; Nature) Dr Jacobs’s team showed that KRAB zinc finger genes (KRAB-ZNFs) in our genome are in a continuous battle against invasions of TEs, revealing how the human genome is actually in a war against itself. His lab currently investigates how the ‘evolutionary arms race’ between TEs and KRAB-ZNFs has re-shaped gene-regulatory networks involved in human brain development. The team find that TE invasions provided our genome with novel gene-regulatory elements, adding an extra level of complexity to how, where and when neuronal genes in our genome are switched-on or -off. In addition, the researchers find that evolutionary changes in KRAB-ZNFs, provide them not only with the capacity to recognise and repress TEs, but also results in them recognising gene promoters. This suggests that both TEs and KRAB-ZNFs have added new primate-specific layers of gene regulation, repeatedly innovating gene expression networks throughout primate evolution. Consistent with this concept, the team presents one of these KRAB-ZNFs which initially evolved to repress TEs, but now has become a modulator of genes central to brain development. The researchers’ work shows that the impact of TE invasions lasts long after the TE has lost its capacity to replicate, which ironically, is largely due to our own genome’s defense mechanism to protect our genome from uncontrolled spreading of TEs.

11:55-12:30 Transposable elements in early mammalian development: do they burst?

Dr Maria-Elena Torres-Padilla, Institute of Epigenetics and Stem Cells, Helmholtz Centre Munich, Germany

Abstract

In mammals, the terminally differentiated sperm and oocyte fuse to create a totipotent zygote upon fertilisation. The mechanisms underlying the epigenetic reprogramming towards totipotency that follows fertilisation are not fully understood, and the molecular features of totipotent cells remain scarce. Embryonic cells remain totipotent only for a restricted time window. During this time, embryonic cells are characterised by an atypical chromatin structure and reactivation of specific families of retrotransposons. Recently, it was reported that totipotent-like cells arise in ES cell cultures in vitro. Like in the embryo, these cells are characterised by the expression of MERVL LTR retrotransposons. To address how the expression of these elements is regulated during the transition between totipotent and pluripotent states, Dr Torres-Padilla’s team first examined histone modifications and chromatin structure in early mouse embryos. Remarkably, the researchers have found that specific features of embryonic chromatin are also present in totipotent-like cells in vitro. Based on this analysis, they have begun to decipher key molecular regulators of repetitive elements in the embryo, and how they contribute to shaping the regulatory programme of the newly formed embryo. The team’s results have identified candidate proteins that regulate chromatin function and expression of these elements and show that they can induce totipotency. The researchers are currently examining the role of these molecules in sustaining totipotency in the embryo. Dr Torres-Padilla will present her team’s latest results that reveal a new role for chromatin integrity in promoting epigenetic reprogramming and sustaining molecular features of totipotent cells.

13:50-14:25 Unmasking transposable elements in regulation and disease

Dr Guillaume Bourque, McGill University, Canada

Abstract

A substantial proportion of the genome of many species is derived from transposable elements (TEs). Moreover, through various self-copying mechanisms, TEs continue to proliferate in the genomes of most species. TEs have contributed numerous regulatory, transcript and protein innovations and have also been linked to disease. However, notwithstanding their demonstrated impact, many genomic studies still exclude them because their repetitive nature results in various analytical complexities. Fortunately, a growing array of methods and software tools are being developed to cater for them. Dr Bourque will present a summary of computational resources for TEs and highlight some of the challenges and remaining gaps to perform comprehensive genomic analyses that do not simply 'mask' repeats. As an example, Dr Bourque will also describe a tool called PopSV that was developed to detect copy number variants (CNVs), which are known to affect a large portion of the human genome and have been implicated in many diseases. Although whole-genome sequencing (WGS) can help identify CNVs, most analytical methods suffer from limited sensitivity and specificity, especially in regions of low mappability. Dr Bourque’s team demonstrate that the PopSV calls are stable across different types of repeat-rich regions and validate the accuracy of our predictions using orthogonal approaches. Applying PopSV to 640 human genomes, they find that low-mappability regions are approximately 5 times more likely to harbor germline CNVs, in stark contrast to the nearly uniform distribution observed for somatic CNVs in 95 cancer genomes.

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14:25-15:00 TEtoolkit: Statistically rigorous software for transposon genomics analysis

Dr Molly Hammell, Cold Spring Harbor Laboratory, USA

Abstract

Transposons form more than half the human genome, and several human diseases have been associated with aberrant activity of transposable elements (TEs) via a variety of mechanisms. Aberrant transposon activity has been shown to induce mutations, alter the regulation of adjacent genes, and produce toxic and/or immunogenic proteins. Despite the importance of TEs in human health and development, reads derived from these regions are frequently ignored in most sequencing data analysis protocols due to the difficulty in properly assigning TE-derived reads to the correct locus of origin. Compromises must often be made between certainty about the exact genomic locations of active TEs and sensitivity to the youngest and most potentially dangerous elements. To address this problem, the Gale-Hammell lab has been systematically creating computational approaches to probabilistically handle reads from repetitive regions in genomics datasets: the TEtoolkit. This has allowed for the identification of new factors contributing to TE silencing as well as new contexts in which TEs are considerably more active than expected. In particular, there is now mounting evidence that TEs contribute to neurodegenerative disease.

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15:00-15:30 Tea

15:30-16:05 A functional ‘gene-like’ annotation of transposable elements using full-length cDNA sequencing

Dr Kaushik Panda, Donald Danforth Plant Science Center, USA

Abstract

Transposable elements (TEs) are more mobile genetic units that have been found to play key regulatory roles in gene expression and impact eukaryotic evolution. Sequencing a new genome of any organism has always been accompanied with annotation of both genes and repeat elements like TEs. But, due to the mobile nature of TEs, there have been constant efforts in discovery of non-reference TEs by resequencing genomes especially with the advent of long read sequencing. However, there is a case to be made to move beyond TE discovery towards transcriptional annotation. It is common for genes to have annotated transcriptional start sites, stop sites and exon-intron boundaries whereas TEs are annotated as blocks of DNA without any transcriptional information. This talk will focus on the loss of information and bioinformatic artefacts that arise because of lack of TE transcriptional annotation. In his research Dr Panda has created a transcript-based functional annotation of Arabidopsis TEs using Nanopore full-length cDNA sequencing in a number of TE activated strains. He will show that characterising TE transcripts improves our investigation of TE RNAs by reducing the complex multi-mapping of short reads. This annotation also enables other genomic studies, for example, understanding how TE integration affects regional mRNA production.

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16:05-16:40 TE driven genome dynamics in a plant lineage over 15 My of evolution

Professor Olivier Panaud, University of Perpignan, France

Abstract

Rice is the staple food crop for more than half of the world’s population. Many factors are now jeopardizing a sustainable production of this cereal, among which, population size increase, environmental changes and reduction of arable land caused by cities expansion. As a consequence, rice has been the focus of intense research over the last decades, which lead to the development of large genomic resources such as several high quality genome assemblies for cultivated rice and several of its wild relatives, genome sequences for 3,000 rice varieties and transcriptomic data for many developmental stages and various physiological stresses. Therefore, the genus Oryza (to which cultivated rice belongs) is now one of the best models to study genome evolution using full scale genomic analyses. We took advantage of these resources to address the question of the impact of transposable elements (TEs) on the structure and evolution of this plant genome. Here, we show that TEs contribute to genomic turn-over at a rate which is much higher than in animal kingdom. We then looked at the transpositional landscape of cultivated Asian rice, taking advantage of the availability of the 3,000 genomes dataset. This resource provides the opportunity to study the impact of TEs at the species level. We show that retrotransposons (the major TE type in plants) are very active in this crop and contribute to genome diversification in agro. Our results suggest that one could tentatively search for TE insertion polymorphisms that may have been adaptive in the recent history of rice cultivation.

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16:40-18:15 Poster session

09:00-09:35 Understanding the potential of TEs to regulate gene expression in mammals

Dr Vasavi Sundaram, EMBL-EBI, UK

Abstract

Transposable Elements (TEs) have now been known to participate in gene expression regulation for several decades. This is an advancement for ‘jumping’ genes which were once thought to be ‘junk’ DNA. However, the extent of TEs’ influence on gene regulation is still being discovered. Understanding TEs and their effect on the breadth of gene expression states and cellular behaviour is central to unmasking the function of TEs. Britten and Davidson postulated the ‘gene-battery’ model that guides our current understanding of TEs in gene expression regulation. TEs represent almost half of human DNA and are capable of regulatory functions from harbouring binding sites for various transcription factors and exhibiting tissue-specific epigenetic activity. Due to the redundancy provided by TEs across the genome, the cellular machinery gains widespread access and control on gene expression across the genome. There are several cases over evolution where TEs underwent exaptation and co-option in gene regulatory landscapes. Together with the ‘gene-battery’ model these cases demonstrate the impact of TEs on gene regulation and occasionally dysregulation of in cells.

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09:35-10:10 Transposon repression or how to hit a moving target

10:10-10:40 Coffee

10:40-11:15 Retroelements, their polydactyl controllers and the specificity of human biology

Professor Didier Trono, École Polytechnique Fédérale de Lausanne, Switzerland

Abstract

KRAB-containing zinc finger proteins (KZFPs) repress the transcriptional activity of transposable elements-embedded regulatory sequences (TEeRS) shortly after embryonic genome activation. Many TEeRS subsequently serve as developmental or tissue-specific modulators, but how KZFPs modulate this process is unknown. We identified two primate-specific KZFP paralogs responsible for repressing partly overlapping sets of evolutionarily recent TEs in human ESCs. These KZFPs are expressed in the developing and adult human brain and control the ability of their targets TEeRS to modulate gene expression in iPSC-derived neurons and brain organoids, notably regulating neurotransmitter profile and preventing the induction of neurotoxic retroviral ENV and of an interferon-like response. These data demonstrate that, rather than just silencing TEs in early embryogenesis, KZFPs keep controlling their transcriptional impact later in development and in adult tissues, and that the evolutionary selection of KZFP genes, rather than reflecting a simple arms race, partakes in the domestication of TEeRS towards the genesis of human-specific transcription networks. Our results also imply that sequence alterations in these KZFP genes or their target TE-derived loci could impact on brain development, function and disease susceptibility.

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11:15-11:50 Transposable elements and the regulation of embryonic cell identity

Dr Michelle Percharde, Imperial College and MRC London Institute of Medical Sciences, UK

Abstract

Much research has focused on the contribution made by single-copy genes and their protein products to the process of early embryogenesis. Conversely, transposable elements (TEs) have been greatly understudied, despite comprising nearly half of mammalian genomes. LINE1 retrotransposons are the most abundant TE class and thought to be largely deleterious for cells due to their ability to ‘copy and paste’ themselves into new sites. However paradoxically, LINE1 is expressed during early mouse embryo development, suggesting a potential function in normal developmental processes. Here Dr Percharde reports that LINE1 plays essential roles in mouse embryonic stem cells (ESCs) and in pre-implantation embryos. In ESCs, LINE1 RNA acts as a nuclear RNA scaffold that recruits Nucleolin and Kap1/Trim28 to repress Dux, a master activator of a transcriptional program specific to the totipotent 2-cell embryo. LINE1 depletion causes inappropriate activation of Dux, along with genes and transcripts driven by the 2-cell specific LTR retrotransposon, MERVL. Intriguingly, this same complex also participates in gene activation, whereby LINE1-dependent association at rDNA repeats promotes rRNA synthesis, hypertranscription and ESC self-renewal. In embryos, LINE1 is required for correct Dux silencing, rRNA synthesis, and exit from the 2-cell stage. These results reveal an essential partnership between LINE1 RNA and chromatin factors in the regulation of transcription, developmental potential and ESC self-renewal.

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11:50-12:25 3'UTR SINE-Directed mRNA Metabolism

Professor Lynne Maquat, University of Rochester, USA

Abstract

Primate-specific Alu SINEs as well as rodent-specific B and ID (B/ID) SINEs can promote mRNA export from the nucleus to the cytoplasm, translation in the cytoplasm and/or Staufen-mediated mRNA decay (SMD) when present in mRNA 3'UTRs. The transposable nature of SINEs, their presence in long noncoding RNAs, their interactions with Staufen (STAU), and their rapid divergence in different evolutionary lineages suggest they could have generated substantial modification of post-transcriptional gene-control networks during mammalian evolution. Some of the variation in SMD regulation produced by SINE insertion might have been similarly adaptive in separate mammalian lineages, leading to parallel evolution of the STAU network by independent exaptation of SINEs. To explore this, Professor Maquat’s team searched for orthologous gene pairs, each carrying a species-specific 3'UTR SINE and each regulated by SMD, by measuring changes in mRNA abundance after individual depletion of two SMD factors, STAU1 and UPF1, in both human and mouse myoblasts. They identified and confirmed orthologous gene pairs with 3'UTR SINEs that have been independently exapted in mouse and humans for SMD control of myoblast metabolism. Their work reveals a novel mechanism for the convergent evolution of post-transcriptional gene regulatory networks in mammals by species-specific SINE transposition and SMD.

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13:35-14:10 Insertion variants at disease risk loci

Dr Kathleen Burns, Johns Hopkins University School of Medicine, USA

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14:10-14:45 Somatic LINE retrotransposition on zebrafish

Dr Jose Luis García-Pérez, MRC Human Genetics Unit, UK

14:45-15:15 Tea

15:15-15:50 The role of natural transposable element insertions in stress response

Dr Josefa González, Institute of Evolutionary Biology, Spain

15:50-16:25 Transposable elements and alternative splicing in cancer

Dr King Jordan, Georgia Institute of Technology, USA

Abstract

Transposable element (TE) derived sequences comprise almost half of the human genome, and their presence has been documented to alter transcripts in a number of different ways, including the generation of alternatively spliced isoforms. Alternative splicing has in turn been associated with tumorigenesis for a number of different cancers. The objective of this study was to broadly characterise the role of human TEs in generating alternatively spliced isoforms in cancer. To do so, Dr King Jordan’s team screened for the presence of TE-derived sequences co-located with alternative splice sites that are differentially utilised in paired normal versus cancer tissues. They analysed a comprehensive set of alternative splice variants from 8,705 matched normal-tumor tissue pairs characterised via RNA-seq as part of The Cancer Genome Atlas (TCGA). Their algorithm uncovered close to 500,000 TE-generated alternative splice events distributed among Catalogue Of Somatic Mutations In Cancer (COSMIC) census genes that have been causally implicated in cancer, and alternative splice sites were significantly enriched within TEs for cancer-implicated genes. SINEs and LINEs were found to contribute the majority of TE-generated alternative splice sites in cancer genes, and TE sequences were equally likely to be involved in the elimination of canonical splice sites or the generation of novel splice sites in cancer tissues. Differential expression analysis was used to detect TE-derived splicing events that are over-expressed in cancer tissues. A number of cancer-associated genes – ATM, MUC16, and MYC – were shown to have overexpressed TE-generated isoforms across a range of cancer types.

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16:25-17:00 Birth of circadian enhancers from transposable elements

Professor Cédric Feschotte, Cornell University, USA

Abstract

Circadian regulators (CRs) are transcription factors that activate or repress circadian gene expression through an oscillatory feed-forward mechanism. The researchers found that ~14% of CR binding sites in the mouse liver are derived from transposable elements and other repeats. While these repeat-associated binding sites (RABS) display comparable oscillatory CR binding to non-RABS, a smaller fraction is characterized by hallmarks of active enhancers. RABS are enriched near genes associated with murine-specific phenotypes. RSINE1, an abundant rodent-specific SINE, contributed a disproportionate amount of RABS. Sequence analyses and reporter assays show that the circadian regulatory propensity of RSINE1 can be explained by the presence of imperfect CR and nuclear receptor binding motifs in the RSINE1 consensus sequence, which have matured after insertion into canonical motifs through recurrent point mutations. Furthermore, CR-bound RSINE1 elements are enriched near evolutionarily older circadian regulatory elements, suggesting that their insertion close to these ancestral elements favored their maturation into redundant enhancers. Together these data point to a model whereby the birth of enhancers from transposons is conditioned not only by the ancestral sequence of the transposon and its post-insertional modification by mutation, but also by the cis-regulatory landscape surrounding the insertion site. The model illuminates how transposition fuels the emergence and turnover of enhancers during mammalian evolution.

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