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Biology of PIWI-Interacting Rnas: New Insights Into Biogenesis and Function Inside and Outside of Germlines

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Biology of PIWI-Interacting Rnas: New Insights Into Biogenesis and Function Inside and Outside of Germlines Downloaded from genesdev.cshlp.org on October 2, 2021 - Published by Cold Spring Harbor Laboratory Press REVIEW Biology of PIWI-interacting RNAs: new insights into biogenesis and function inside and outside of germlines Hirotsugu Ishizu,1,2 Haruhiko Siomi,2 and Mikiko C. Siomi1,2,3,4 1Graduate School of Science, The University of Tokyo, Tokyo 113-0033, Japan; 2Keio University School of Medicine, Tokyo 160-8582, Japan; 3Core Research for Evolutional Science and Technology (CREST), Japan Science and Technology Agency (JST), Saitama 332-0012, Japan PIWI-interacting RNAs (piRNAs) are endogenous small RNAs (piRNAs). These small RNAs can be classified on noncoding RNAs that act as guardians of the genome, the basis of their origins, processing factors, and Argo- protecting it from invasive transposable elements in the naute-binding partners (Kim et al. 2009). piRNAs are germline. Animals lacking piRNA functions show defects generated from single-stranded precursors in a manner in gametogenesis and exhibit sterility. Their descendants independent of RNase III enzymes (Vagin et al. 2006; are also predisposed to inheriting mutations. Thus, the Brennecke et al. 2007; Houwing et al. 2007), which are, in piRNA pathway has evolved to repress transposons post- contrast, necessary for endo-siRNA and miRNA biogen- transcriptionally and/or transcriptionally. A growing num- esis. piRNAs associate with PIWI subfamily members of ber of studies on piRNAs have investigated piRNA-medi- the Argonaute family of proteins, while endo-siRNAs and ated gene silencing, including piRNA biogenesis. However, miRNAs associate with AGO subfamily members. piRNAs piRNAs remain the most enigmatic among all of the are normally 24–32 nucleotides (nt) long, but endo-siRNAs silencing-inducing small RNAs because of their com- and miRNAs are 20–23 nt in most cases. Thus, the plexity and uniqueness. Although piRNAs have been uniqueness of piRNAs is obvious. previously suggested to be germline-specific, recent stud- piRNAs arise from intergenic repetitive elements in ies have shown that piRNAs also play crucial roles in the genome called piRNA clusters (Brennecke et al. 2007). nongonadal cells. Furthermore, piRNAs have also recently piRNA clusters span a wide region of the genome, occa- been shown to have roles in multigenerational epigenetic sionally consisting of >100,000 bases, and are mostly phenomena in worms. The purpose of this review is to comprised of various transposable DNA elements and their highlight new piRNA factors and novel insights in the remnants. Thus, piRNAs, especially those in Drosophila piRNA world. gonads and mouse prepachytene piRNAs (piRNAs ex- pressed before the pachytene stage of meiosis in spermato- genesis), are enriched in transposon sequences. Most of RNA silencing controls gene expression in a spatiotem- these piRNAs have an antisense orientation to trans- poral-specific manner to tightly regulate the development poson transcripts and hence can induce silencing by and homeostasis of living bodies (Bartel 2009; Malone and hybridizing with them (Saito and Siomi 2010). Loss-of- Hannon 2009). The key components of RNA silencing— function mutations in piRNAs and their cofactors, PIWI small noncoding RNAs and Argonaute proteins—associate proteins, derepress transposons, allowing them to insert stoichiometrically to form RNA-induced silencing com- copies of themselves or relocate within the genome in a plexes (RISCs), the effector complexes in RNA silencing random fashion (Kalmykova et al. 2005). This selfish event (Siomi and Siomi 2009). RISCs are directed to the target activates the Chk2 DNA damage checkpoint, resulting in genes based on the complementarities between small defects of, for example, microtubule organization and axis RNAs and target gene transcripts and inhibit their expres- specification during gonadal development, which often sion by cleaving the transcripts or inducing translational lead to infertility (Khurana and Theurkauf 2010). inhibition, RNA instability, and/or heterochromatiniza- The unique characteristics of piRNAs have attracted tion (Moazed 2009; Siomi et al. 2011). many researchers, encouraging them to unveil the Animal species express three types of endogenous mystery of piRNAs. Remarkable progress has been silencing-inducing small RNAs: microRNAs (miRNAs), made, especially in the area of biogenesis. A compre- endogenous siRNAs (endo-siRNAs), and PIWI-interacting hensive computational analysis of piRNA populations in the gonads of various animals—both wild type and [Keywords: PIWI; germline; piRNA] mutants showing defects in oogenesis and/or spermato- 4Corresponding author E-mail [email protected] genesis—led to two models for piRNA biogenesis: Article is online at http://www.genesdev.org/cgi/doi/10.1101/gad.203786.112. the primary processing pathway and the amplification GENES & DEVELOPMENT 26:2361–2373 Ó 2012 by Cold Spring Harbor Laboratory Press ISSN 0890-9369/12; www.genesdev.org 2361 Downloaded from genesdev.cshlp.org on October 2, 2021 - Published by Cold Spring Harbor Laboratory Press Ishizu et al. loop (see below, ‘‘Outline of the piRNA Biogenesis The amplification loop Pathway’’). Primary piRNAs are subjected to an amplification system However, the mechanisms underlying piRNA biogen- to enforce the high expression of piRNAs in the germline: esis and functions remain largely unknown, mainly This system is called the amplification loop or the Ping- because the piRNA pathway has little in common with Pong cycle (Fig. 1; Brennecke et al. 2007; Gunawardane the endo-siRNA and miRNA pathways as well as the et al. 2007; Houwing et al. 2008). In this system, Aub in restriction of piRNA territories to the reproductive flies and MILI in mice associated with primary piRNAs tissues. Nonetheless, recent studies have suggested a pos- cleave the target RNAs via their Slicer (endonuclease) sible function of piRNAs in nongonadal cells such as activity (Saito et al. 2006; Gunawardane et al. 2007; neuronal cells in Aplysia and mammals (Lee et al. 2011; Nishida et al. 2007). This process forms the 59 ends of Rajasethupathy et al. 2012). Other studies have charac- secondary piRNAs. In Drosophila, maternally deposited terized new piRNA factors, Vreteno (Vret) and Shutdown piRNAs can also trigger the Ping-Pong cycle, as do (Shu) (Handler et al. 2011; Zamparini et al. 2011; Olivieri primary piRNAs (Brennecke et al. 2008). The cleavage et al. 2012; Preall et al. 2012; Xiol et al. 2012), and revealed products are then transferred onto other PIWI members, Zucchini (Zuc) as an endonuclease necessary for primary Ago3 in flies and MIWI2 in mice, and are trimmed from piRNA biogenesis (Ipsaro et al. 2012; Nishimasu et al. the 39 end to give rise to mature piRNAs. This step is 2012). Multigenerational epigenetic phenomena in nem- atodes are now known to involve piRNAs (Ashe et al. mechanistically equivalent to that observed in the pri- 2012; Shirayama et al. 2012). Thus, the visibility of the mary processing pathway (Fig. 1), although the factors piRNA field has rapidly broadened. required might differ. In turn, in flies, Ago3 associated with secondary piRNAs cleaves its targets, giving rise to secondary piRNAs, which associate with Aub (Brennecke Outline of the piRNA biogenesis pathway et al. 2007; Gunawardane et al. 2007; Li et al. 2009; Malone et al. 2009). Through these reciprocal Slicer- The primary processing pathway mediated cleavage reactions between Aub and Ago3, the Nascent transcripts arising from piRNA clusters are heterotypic Ping-Pong cycle persists in Drosophila processed into piRNA-like molecules, which are then (Fig. 1). Piwi is translocated to the nucleus upon primary loaded onto PIWI proteins (Fig. 1). The factors involved in piRNA loading (Haase et al. 2010; Olivieri et al. 2010; the transcription of piRNA clusters and its regulation Saito et al. 2010; Ishizu et al. 2011); therefore, Piwi barely remain elusive. The current model proposes that the 59 contributes to secondary piRNA production via the am- ends of piRNAs are determined prior to loading onto PIWI plification loop (Malone et al. 2009; Olivieri et al. 2012). proteins. However, the 39 ends of piRNAs harbor extra In the Drosophila gonadal soma, Piwi is expressed and bases, which need to be trimmed upon association with associates with primary piRNAs. However, the expres- PIWI proteins (Kawaoka et al. 2011; Vourekas et al. 2012). sion levels of Aub and Ago3 are too low for them to The lengths of mature piRNAs are determined during operate in the amplification loop. Thus, the situation in this step, largely depending on the sizes of the PIWI ovarian somatic cells, such as follicle cells, is different proteins. Thus, piRNAs associated with individual PIWIs from that in germ cells. show a similar but distinct size distribution (Siomi et al. Like Piwi in Drosophila, in mice, MIWI2 loaded with 2011). The factors responsible for the 39 trimming secondary piRNAs is localized to the nucleus upon remain elusive. Upon maturation, the 39 ends of piRNAs piRNA loading (Fig. 1; Carmell et al. 2007). Thus, MIWI2 are 29-O-methylated by Hen1/Pimet, which is associated likely does not contribute to the bearing of secondary with PIWI proteins (Horwich et al. 2007; Saito et al. 2007). piRNAs via the Ping-Pong cycle. The pathway for pro- This modification maintains the stability of piRNAs in ducing MIWI2–piRNAs is thus considered to be the ‘‘one- vivo (Horwich et al. 2007; Kamminga et al. 2010, 2012; way’’ secondary piRNA biogenesis pathway (Aravin et al. Billi et al. 2012). Hen1 was first discovered in Arabidopsis 2008; De Fazio et al. 2011; Reuter et al. 2011), although as the enzyme responsible for 29-O-methylation of miRNAs there is evidence supporting the notion that the MILI– (Chen 2005). However, in animals, Hen1 does not 29-O- piRNA complex may operate a homotypic Ping-Pong methylate miRNAs. Although there is no direct evidence cycle in mouse testes (De Fazio et al. 2011). MIWI is for this, the high turnover rate for animals’ miRNAs may associated with pachytene piRNAs (piRNAs expressed serve to spatiotemporally regulate their function.
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