ARTICLE IN PRESS Review Endocrinol Metab 2020 Forthcoming. https://doi.org/10.3803/EnM.2020.302 Article pISSN 2093-596X · eISSN 2093-5978 WD40-Repeat Proteins in Ciliopathies and Congenital Disorders of Endocrine System Yeonjoo Kim, Soo-Hyun Kim Cell Biology Research Centre, Molecular and Clinical Sciences Research Institute, St. George’s, University of London, London, UK WD40-repeat (WDR)-containing proteins constitute an evolutionarily conserved large protein family with a broad range of biologi- cal functions. In human proteome, WDR makes up one of the most abundant protein-protein interaction domains. Members of the WDR protein family play important roles in nearly all major cellular signalling pathways. Mutations of WDR proteins have been as- sociated with various human pathologies including neurological disorders, cancer, obesity, ciliopathies and endocrine disorders. This review provides an updated overview of the biological functions of WDR proteins and their mutations found in congenital disorders. We also highlight the significant role of WDR proteins in ciliopathies and endocrine disorders. The new insights may help develop therapeutic approaches targeting WDR motifs. Keywords: WDR proteins; Ciliopathies; Congenital, hereditary, and neonatal diseases and abnormalities; Neuroendocrine; Kall- mann syndrome INTRODUCTION proteins (based on UniProt) discussed in this review are sum- marised in the Supplemental Table S1. WDR proteins associated WD40-repeat (WDR) refers to a series of loosely conserved with pathological conditions that are not discussed in the main structural motifs comprised of approximately 40 amino acids, text in detail are summarised in the Supplemental Table S2. often terminating in tryptophan (W)-aspartic acid (D). WDR protein family is a large group of proteins commonly possessing MOLECULAR STRUCTURE OF WDR the WDR motifs, that are involved in a wide range of important PROTEINS biological processes. Inherited or acquired defects in WDR pro- WDR motif was first described in the β-subunit of a GTP-bind- teins result in numerous health problems including neurological ing protein transducin complex as a sequence of repeats of 40 to diseases, ciliopathies, and cancers. In this review, we provide a 60 amino acids that begin with glycine and histidine (GH) and unique overview and discussion on the molecular mechanisms end with tryptophan and aspartic acid (WD) dipeptides [1]. and functions of WDR proteins, especially focusing on those WDR is an evolutionarily conserved and highly abundant do- that have been associated with human congenital disorders and main in eukaryotes with nearly 1% of human proteomes con- endocrine diseases. Many of the WDR proteins are called differ- sisting of WDR-containing proteins [2]. Most recent protein do- ent names mainly due to historical reasons. The official gene main database (SMART, http://smart.embl.de/) predicts that 921 nomenclature along with full and alternative names of WDR WDR proteins are encoded in humans, 591 in Mus musculus Received: 9 July 2020, Revised: 3 August 2020, Accepted: 10 August 2020 Copyright © 2020 Korean Endocrine Society Corresponding author: Soo-Hyun Kim This is an Open Access article distributed under the terms of the Creative Com- Cell Biology Research Centre, Molecular and Clinical Sciences Research mons Attribution Non-Commercial License (https://creativecommons.org/ Institute, St. George’s, University of London, Cranmer Terrace, London SW17 licenses/by-nc/4.0/) which permits unrestricted non-commercial use, distribu- 0RE, UK tion, and reproduction in any medium, provided the original work is properly Tel: +44-208-266-6198, E-mail: [email protected] cited. www.e-enm.org page 1 of 13 ARTICLE IN PRESS Kim Y, et al. and 212 in Drosophila melanogaster. WDR proteins are rarely WDR PROTEINS IN HUMAN DISEASE present in prokaryotes [2]. Each WDR protein can have 4 to 16 copies of WDRs forming seven or more bladed beta-propeller According to the Online Mendelian Inheritance in Man folds [3], which can provide three structural surfaces (top, side, (OMIM) database (https://omim.org/), there is a significant cor- and bottom region of propeller) available to interact with other relation between WDR protein dysfunction and human diseases binding partners [2]. Based on these structural features, it is sug- (Fig. 1). Among 360 WDR genes we assessed, 79 genes were gested that WDR proteins could serve as a scaffold that medi- reported to be associated with human pathologies which include ates protein-protein or protein-DNA interaction [4]. Since neurological disorders (40.5%), ciliopathies (21.5%), immune WDRs do not possess any catalytic activity themselves, func- diseases (8.9%), eye problems (7.6%), skeletal anomalies tional diversity is likely achieved by coordination of multiple (3.8%), cancers (3.8%), endocrine disorders (2.5%), inflamma- binding partners. Mutations in WDR proteins have been report- tion (2.5%), and others including preimplantation embryonic le- ed in several human diseases. Notably, clinically identified mu- thality (8.9%). tations of WDR are mostly found on the surface of the protein, Notably, a significant number of WDR proteins have been as- presumably interfering their binding interactions with other pro- sociated with ciliopathies, a group of genetic disorders resulting teins [1,2,4]. from defects in the structure or function of cilia. Cilia are highly conserved microtubule-based hair-like organelles that extend BIOLOGICAL FUNCTIONS OF WDR from the plasma membrane of most vertebrate cells. Cilia can PROTEINS broadly be classified into two types, motile and non-motile (pri- mary) cilia that share the principal axoneme structures [30,31]. WDR proteins can be primarily defined by their sequence simi- The axoneme consists of a circular arrangement of nine pairs of larity in the WD40-repeat domain. However, a wide range of microtubules called outer doublets. In addition to the outer dou- sequence variation has been found in WD40-repeats, resulting blets, motile cilia contain a pair of microtubules in the centre in variable numbers of beta-propeller structures. Variations out- called inner doublets [32]. This central pair of microtubules is side of the WD-repeat domain can also contribute to the multi- the scaffold of the central pair complex including radial spokes, domain contexts [5]. In fact, although all WDR proteins are inner and outer dynein arms and nexin links [33]. The intrafla- structurally related, their molecular functions can be quite dis- gellar transport (IFT) particles assemble and maintain the cilium tinct. This functional diversity is usually acquired from the ad- by trafficking ciliary proteins within the cilium [22,23]. Two ditional domains present in the respective WDR proteins. Cur- rently, more than 360 additional domains are reported in WDR 2.5 2.5 3.8 proteins [6]. The most commonly found additional domains are 3.8 shown to be functionally involved with ubiquitylation (e.g., F- Neurological disorders Ciliopathies box [7], SOCS-box [8], RING-finger [9]), microtubule dynam- 7.6 ics (e.g., CTLH domain [10]), phospholipid-binding (e.g., Others 40.5 Immune diseases FYVE domain [11]) and endocytic vesicle coating (e.g., clathrin 8.9 Eye problems terminal domain [12]). WDR domains are also identified in es- Skeletal anomalies sential subunits of multiprotein complexes that participate in 8.9 Cancers various signalling pathways regulating DNA repair [13-15], 21.5 Endocrine disorders Inflammation protein degradation [16,17], cell cycle control [18,19], mRNA translation [20,21], cilia assembly and maintenance [22,23], and hormone biosynthesis [24]. Therefore, it is not surprising that Fig. 1. A chart indicating the relevant prevalence of human diseases WDR proteins play important roles in fundamental cellular associated with WD40-repeat (WDR) proteins. The data are based functions, such as signal transduction, gene expression, RNA on the entries in Online Mendelian Inheritance in Man (OMIM). Total 79 out of 360 WDR proteins have been linked with disease processing, protein synthesis, homeostasis, proliferation, apop- categories as indicated. The ‘others’ category includes multi-organ tosis, intracellular vesicle trafficking, and cargo recognition defects, liver failure, cardiovascular defects and embryonic lethali- [2,25-29]. ty. The full list of WDR proteins assessed are included in Supple- mentary Table S1. page 2 of 13 www.e-enm.org Endocrinol Metab 2020 Forthcoming. Posted online 2020 ARTICLE IN PRESS WDR Mutations in Human Diseases subcomplexes IFT-A and IFT-B, consisting of at least 6 and 13 Mutations in LRRK2 are the most common cause of autoso- proteins, respectively, move along the cilium bidirectionally via mal dominant Parkinson’s disease [60,61]. To date, more than a retrograde (IFT-A) and anterograde (IFT-B) transport. In retro- hundred mutations of LRRK2 have been identified and six of grade transport (from the ciliary tip to the base), IFT-A uses dy- them are confirmed to be pathogenic [62]. LRRK2 protein binds nein-2 as a motor, whereas IFT-B is powered by kinesin-2 for to the synaptic vesicles and regulates vesicular trafficking by in- anterograde movement [34,35]. Many IFT proteins contain pro- teracting with pre-synaptic proteins such as actin and synapsin tein-protein interaction motifs including WDRs, tetratricopep- [63]. The sequence variation of glycine to arginine at residue tide repeats and coiled coils motifs [22], facilitating the interac- 238 (G2385R) which is located between the 5th