Evolutionarily Conserved Sperm Factors, DCST1 and DCST2, Are Required for Gamete Fusion Naokazu Inoue1*, Yoshihisa Hagihara2, Ikuo Wada1
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SHORT REPORT Evolutionarily conserved sperm factors, DCST1 and DCST2, are required for gamete fusion Naokazu Inoue1*, Yoshihisa Hagihara2, Ikuo Wada1 1Department of Cell Science, Institute of Biomedical Sciences, School of Medicine, Fukushima Medical University, Fukushima, Japan; 2Biomedical Research Institute, National Institute of Advanced Industrial Science and Technology (AIST), Ikeda, Japan Abstract To trigger gamete fusion, spermatozoa need to activate the molecular machinery in which sperm IZUMO1 and oocyte JUNO (IZUMO1R) interaction plays a critical role in mammals. Although a set of factors involved in this process has recently been identified, no common factor that can function in both vertebrates and invertebrates has yet been reported. Here, we first demonstrate that the evolutionarily conserved factors dendrocyte expressed seven transmembrane protein domain-containing 1 (DCST1) and dendrocyte expressed seven transmembrane protein domain-containing 2 (DCST2) are essential for sperm–egg fusion in mice, as proven by gene disruption and complementation experiments. We also found that the protein stability of another gamete fusion-related sperm factor, SPACA6, is differently regulated by DCST1/2 and IZUMO1. Thus, we suggest that spermatozoa ensure proper fertilization in mammals by integrating various molecular pathways, including an evolutionarily conserved system that has developed as a result of nearly one billion years of evolution. *For correspondence: [email protected] Introduction Gamete recognition and fusion in mammals are considered to occur through a complex intermolecu- Competing interests: The lar interaction in which izumo sperm–egg fusion 1 (IZUMO1), sperm acrosome associated 6 authors declare that no (SPACA6), transmembrane protein 95 (TMEM95), fertilization influencing membrane protein (FIMP) competing interests exist. and sperm–oocyte fusion required 1 (SOF1) on the sperm side and JUNO (also known as IZUMO1 Funding: See page 10 receptor) and cluster of differentiation 9 (CD9) on the ovum side are all involved in membrane fusion, Received: 07 January 2021 as proven by gene disruption (Barbaux et al., 2020; Bianchi et al., 2014; Fujihara et al., 2020; Accepted: 23 March 2021 Inoue et al., 2005; Kaji et al., 2000; Lamas-Toranzo et al., 2020; Le Naour et al., 2000; Published: 19 April 2021 Lorenzetti et al., 2014; Miyado et al., 2000; Noda et al., 2020). Particularly, the IZUMO1–JUNO interaction is likely to be essential for triggering gamete fusion (Aydin et al., 2016; Inoue et al., Reviewing editor: Steven W 2015; Inoue et al., 2013; Ohto et al., 2016); however, it is unknown how these factors contribute L’Hernault, Department of Biology, Emory University, to this process (Bianchi and Wright, 2020). Atlanta, Georgia Here, we show that an osteoclast fusion-related factor, dendrocyte expressed seven transmem- brane protein (DC-STAMP) (Kukita et al., 2004; Yagi et al., 2005), homologue DC-STAMP domain- Copyright Inoue et al. This containing 1 (DCST1), and its paralogue DCST2, which show testis and haploid-specific expression article is distributed under the (Figure 1D), are indispensable for fertilization in mice. Mouse DCST1 is considered to be an ortho- terms of the Creative Commons Attribution License, which logue of Caenorhabditis elegans spermatogenesis-defective 49 (SPE-49) (Wilson et al., 2018) and permits unrestricted use and Drosophila SNEAKY (Wilson et al., 2006), whereas mouse DCST2 is considered to be an orthologue redistribution provided that the of C. elegans SPE-42 (Kroft et al., 2005) and Drosophila DCST2, although the numbers of putative original author and source are transmembrane regions appear different (Figure 1A,B). Previously, these factors have been shown credited. to be essential for fertilization. In fact, phylogeny analysis revealed that DCST1 and DCST2 belong Inoue et al. eLife 2021;10:e66313. DOI: https://doi.org/10.7554/eLife.66313 1 of 12 Short report Developmental Biology A DCST1 human 1 M----------------------------DIKHHQNGTRGQRRKQPHTTVQRLLTWGLPVSCSWFLWRQPGEFPVTALLLGAGAGGLLAIGLFQLLVNPMNIYEEQKIMFLYSLVGLGAMGWGTSPHIRCASLLLVPKMLGKEGRLFVLG 122 mouse 1 MAFLSSTLHSLGIFEKISRIKEVLKNRLLDLTKRRDQAREQQRKRPHTIIQGLLLWSLPVSWIRFLWRQPGEFPVTAFLLGAGTGGLLAIGLFQLLVNPMNIYEEQKVVALYCLASLGAIGWGTSPHIRCASLLLVPKMLGKEGRLFVMG 150 nematode 1 MGFFDD--------KKRDEVRRRENIG--FLTDYLVRSEVSDYYRLRLLVNFLFGLLISSSLYHAGWK------NLNFGDFDYTYGL---------------------IVKWTIVAFSTYAFAVSPIFRCALFCVLIGAFGKQGQYPFTM 113 fruit fly 1 MFHFLT--------KSIRQAIGRLACRPLYCLLYGQESEDEPWLKP---VRYVARLLVGLFLAYLLWG----LVALNFNLGRWLAPL-----------------PVHVPVFEVFVLLNGLAFMLIRSVRAVTLLMFVALVGKSGRSYLRA 118 * . .* . .. .* . .. *.. .. ** *. human 123 YALAAIYVGPVANLRHNLNNVIASLGCTVELQINNTRAAWRISTAPLRAMFKDLLS----SKELLRAETRNISATFEDL--------DAQVNSETGYTPE-------------DTMDSGETAQGREARQ-APASRL--HLSTQK--MYEL 242 mouse 151 YALAAIYSGPAANLRSNINEVIASLGCTVELQINNTRSAWRVSTAPLRAVFKGMVG----SKDSLNKEIQNVSTSFEEM--------DEQVKSDAGYSSE-------------------DWDKNRESTEMFGTSRVRPYLSTQM--VYEL 267 nematode 114 LVMSNLQEGPISNMMSNYETTSEIVMCHIELQSKITANRVALLTGPLEELIEKLMAKGIRAMKAVSRETRALITPFMELLKTEKTKIDKKIEIERGQLEDIEKRKQRILKMWEKSMNRSLTEEDQIAGELLPAEDLIDNVTIGEPPVWKK 263 fruit fly 119 IAFAFVIAGPIDNLASNAGEVARVFACTTVLTYNLTKTRFDLMARPFTSTLQHMRG-----------DIAEIRNTFREL---------EQVLGELKYAVE--------------------QESHDEDHTTAPQSELPSADAVRE--NFLR 226 . ** *. .* . *. .*. * . *. * .. .. .. human 243 KTKLRCSYVVNQAILSCRRWFDRKHEQCMKHIWVPLLTH---LLCLPMKFKFFCGIAKVMEV--WCRNRIPVEGNFGQTYDSLNQS------IRGLDGEFSANIDFKEEKQAGVLGLN-TSWERVSTEVR--DY---VYRQEARLEWALG 375 mouse 268 RTRLRCIHVVNKAILSCYRWFDKKHKNCMRRIHLPLFNN---MICVPMKFKFLCNIAKVIEI--WCYKRIPVEGNFGQTYDSVNQS------IHGLSGEFSANINLKEEKQSSMVGLNTTNWEHMGTEVR--DY---VRQQETYLQWAMG 401 nematode 264 FKTPLIQKMADKMSRNCEQMFNKGIDKC-RNLAAELVSSCKDAIIWPIE-AFIC---PKLNVEGVCDAVERRVQSMNICRNQLEESHVDPSIEKDVTDVMKLTEELEQNSNIELHSVR-VETPRVAIEYRLSDLKIKIRAANLYFKSIVG 407 fruit fly 227 NMQHRCKQQLRSGTRVCQEVFRKGYQKCSTNFPDFLAA----AICWPYRVDIICRLNPFGSQDKICDASEVVPQNFGQAYVDMLRT------ERDLFDA-SADIEISYELRNETVKGQ-LLTAQQTSEAFMKDF---NRRRRIFTTTM-- 359 .. * * . * * .. * . * * . .... .. * . * . human 376 LLHVLLS---CTFL-LVLHASFSYMDSYNHDIRFDNIYISTYFCQIDDRRKKLGKRTLLPLRKAEEK---TVIFPCKPTIQASEMSNVVRELLETLPILLLLVVLCGLDWALYSIFDTIRHHSFLQYSFRSSHKLEVKVGGDSMLARLLR 518 mouse 402 LLHVLLS---CTFL-LVFHSAFSYMDHYNWDIRFDNIYISTYFCQIDARRKKLGKQSLLPLRKAERK---TVIFPFKATIQAWEMRYVIRELLETLPIVLLLLVLCAIDWALYSVFDTIRQHSFVQYSFRSSHKLEIKVEGDSILAKLLR 544 nematode 408 VSKQIFQAFFIYFVYTIFRDSVGMIRKYQEDVAFSNSFVTKEFWMIDRFRQARGQTHLSHFSKQEKREW-RVMEVFSLPTKA-ERSKAVRPFFRWFILALTVSVIIILDYYLFVFLDSVVESARQQVKQKA------------------- 536 fruit fly 360 ---QILEKFLCLFLLRVIYASIGYYFQYRGNVEFDNFYITDYFKHIDAERKANEKRSILPLHSHERANYVDVAHVCSRTAK--ESLTAMYHLLQLSLEVITAGLFLLLDRMVVGLLQIIRQRSLITYQQEGEHEIRFHINGTGLMARLLR 504 . *. * . *.* .. .* ** *. .. .... * * . * . .. .* . .. .. ... human 519 KTIGALNTSSETVMESNNMPCLPQPVGLDAR----AYWRAA---------------------------------VPIGLLVCLCLLQAFGYRLRRVIAAFY----FPKREKKRILFLYNDLLKKR-AAFTKLRRAAILRRERQQKAPRHP 626 mouse 545 KTIGALNTSSSTDVETNNMPCLPQPISLNAR----DYFKAS---------------------------------LPTLLLVCLCLAQAFGYRLRRVIAAFY----FPKREKKRALFFYNEFLKKR-SAFTKLRRAAIVRRANQQKAPPHY 652 nematode 537 --------------------SAPAGLNITGEGVLADFLKTMTSTNETLEVDQTLSNEHCLLKPLAPNTDILIYWLVIPLLLSFLFQVVFSFAIRRIVLNYFLPFMFPMRSKVRLIQFYNKCLVNR----EKHRKEA---RARIRFMVDRR 659 fruit fly 505 TTMRNFNMHERVSTSLSNEECLPNAHALPQR----FYYRLM---------------------------------LLYLAIVLLIYQSTTFLRLRRVICSYF----YDKREKQRVLFLYNSLLRNRLSLFEILRHNAEQNLAARHVQDNLN 613 . ..* . ... ..**.. ...*.* * .. ** * * . ..* * .. human 627 LADILHRGCPLLRRWL------CRRCVVCQAPETPESYVCRTLDCEAVYCWSCWDDMRQRCPVC------TPREEL-------SSSAFSDSNDDTAYAG----------------------------------------------- 706 mouse 653 LVEALYRRCPLLHRFM------RQRCVVCQAMETPDSYVCPTPDCKALYCRSCWDDMQRLCPVC------TPREEL-------SSSAHSDSNDDAVYGD----------------------------------------------- 732 nematode 660 RIETLRKGEFSAGGSF------LKQHVFERIFKTAK---------------------------CFLCQEKTYARKLFYCAEVKCQSSFCKDCVEDNFGKCYSCMFIKKEVTDQKSKLLNPDEVKTKDQFLSYGEFLERIQQDHMQK 772 fruit fly 614 IVLRLRLLCPGVFGCLQHLSCGKRNCIICDALEDPDFVLCS--GCGLPYCHECSVMLKNVCIKCGLAISFQPFTML--------DEAWSDDSSEELYDF-------------------------------------------RKDK 706 * .. .. ... .. * .. * .. .. DCST2 human 1 MPKVMKDVVHPLGG--EEPSMARAVVRSVGGFTL----------GLSLATAYG-----LLELLVEGHSPWG----------------------------CLVGTLTLAAFLSLGMGFSR-------------QVRATVLLLLPQAF---- 88 mouse 1 ------------ME--EGSSTARAVVRSTCGFTV----------GLSLATAFG-----LLELLGEGHSPFG----------------------------CLVTTVTLAAFLSLGMGFSR-------------QVRVSVLLLLPQAF---- 76 nematode 1 ------------MGLIGLWNVKRRANRAVRKFQFVVNNIRREKSGQNRVLRYSERLKKWLSVL---YPPWKKEKDDGIYDNLFEDGTRANSIVLLFRNILMIELFGIVMYFLLSFKYSQYPEFIGTSISTIIHVFLIFTLIFPKMMTYFM 135 fruit fly 1 -----MEESEPRYG--AMWTFKLVAPYVIGGYLC----------GLAVTLAWN-----WWQL---GHLVLG-----------------------------LNLTFAIPVALVLAVFYSR-------------PVRCIVTLAVPSLC---- 79 . .. *. .. .. * *. *. * * human 89 -------SRQGRTLLLVAAFGLVLQGPCANTLRNFTRASEAVACGAELALNQTAEVLQRAKQPLVSALNKIKAIARKTKEVADRVRKFFRSIMDGVKHIARALRNVWQWLLHIGDVCNSELGNPYLKCARVFDDAKDSCMMVIPQA---- 227 mouse 77 -------SKQSRLLLLVASFGLVLQGPCANTLQNFTRASEAVACGAELALNQTAEMLERAKQPLISALSKIKAIAQKAKVVGDRIRKFFRSIIDAVKHIARCLQNVWYWLLHIGDMCNSELGNPYSKCTQVFDDAKIHCMKVVSGF---- 215 nematode 136 ISIHVMLTTKIKNIILILLVTMAFEGPAMNVIGNIHQVASGVACVQIDVMSSRNDVEGNVMDKGAMLVSRFRALLQNVSGPMNKVKKMLLVLDEKMTKFVDIMRRHYRVIANLSNQCRNMLKTPYTKCLDIFDDAYFYCKGKARFFGSHG 285 fruit fly 80 -------SSRGRAFLISLAFVIAAVGPTANILANLKVMLRSLACGQELLRQALGQMLDVILEPVNAIQLAVDLLMREVRRVLNLAMVVLLRIQDHLIAIIKTLKNCAAWLKSIVDLCNTEMGTPWARCKKTAHHAMIRCQAKMGVF---- 218 . .. ** .* .