{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2025,11,4]],"date-time":"2025-11-04T23:44:20Z","timestamp":1762299860192,"version":"build-2065373602"},"reference-count":31,"publisher":"MDPI AG","issue":"15","license":[{"start":{"date-parts":[[2021,7,26]],"date-time":"2021-07-26T00:00:00Z","timestamp":1627257600000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"funder":[{"DOI":"10.13039\/501100001809","name":"National Natural Science Foundation of China","doi-asserted-by":"publisher","award":["81971964","31730096"],"award-info":[{"award-number":["81971964","31730096"]}],"id":[{"id":"10.13039\/501100001809","id-type":"DOI","asserted-by":"publisher"}]}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":["IJMS"],"abstract":"<jats:p>Toxoplasma gondii (T. gondii) is an important human and veterinary pathogen causing life-threatening disease in immunocompromised patients. The UBL-UBA shuttle protein family are important components of the ubiquitin\u2013proteasome system. Here, we identified a novel UBL-UBA shuttle protein DSK2b that is charactered by an N-terminal ubiquitin-like domain (UBL) and a C-terminal ubiquitin-associated domain (UBA). DSK2b was localized in the cytoplasm and nucleus. The deletion of dsk2b did not affect the degradation of ubiquitinated proteins, parasite growth in vitro or virulence in mice. The double-gene knockout of dsk2b and its paralogs dsk2a (\u0394\u0394dsk2adsk2b) results in a significant accumulation of ubiquitinated proteins and the asynchronous division of T. gondii. The growth of \u0394\u0394dsk2adsk2b was significantly inhibited in vitro, while virulence in mice was not attenuated. In addition, autophagy occurred in the \u0394\u0394dsk2adsk2b, which was speculated to degrade the accumulated ubiquitinated proteins in the parasites. Overall, DSK2b is a novel UBL-UBA shuttle protein contributing to the degradation of ubiquitinated proteins and is important for the synchronous cell division of T. gondii.<\/jats:p>","DOI":"10.3390\/ijms22157943","type":"journal-article","created":{"date-parts":[[2021,7,26]],"date-time":"2021-07-26T09:25:52Z","timestamp":1627291552000},"page":"7943","update-policy":"https:\/\/doi.org\/10.3390\/mdpi_crossmark_policy","source":"Crossref","is-referenced-by-count":3,"title":["Toxoplasma gondii UBL-UBA Shuttle Protein DSK2s Are Important for Parasite Intracellular Replication"],"prefix":"10.3390","volume":"22","author":[{"given":"Heng","family":"Zhang","sequence":"first","affiliation":[{"name":"National Animal Protozoa Laboratory, College of Veterinary Medicine, China Agricultural University, Beijing 100193, China"},{"name":"Key Laboratory of Animal Epidemiology of the Ministry of Agriculture, College of Veterinary Medicine, China Agricultural University, Beijing 100193, China"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Xu","family":"Yang","sequence":"additional","affiliation":[{"name":"National Animal Protozoa Laboratory, College of Veterinary Medicine, China Agricultural University, Beijing 100193, China"},{"name":"Key Laboratory of Animal Epidemiology of the Ministry of Agriculture, College of Veterinary Medicine, China Agricultural University, Beijing 100193, China"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-9986-8314","authenticated-orcid":false,"given":"Zhu","family":"Ying","sequence":"additional","affiliation":[{"name":"National Animal Protozoa Laboratory, College of Veterinary Medicine, China Agricultural University, Beijing 100193, China"},{"name":"Key Laboratory of Animal Epidemiology of the Ministry of Agriculture, College of Veterinary Medicine, China Agricultural University, Beijing 100193, China"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Jing","family":"Liu","sequence":"additional","affiliation":[{"name":"National Animal Protozoa Laboratory, College of Veterinary Medicine, China Agricultural University, Beijing 100193, China"},{"name":"Key Laboratory of Animal Epidemiology of the Ministry of Agriculture, College of Veterinary Medicine, China Agricultural University, Beijing 100193, China"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Qun","family":"Liu","sequence":"additional","affiliation":[{"name":"National Animal Protozoa Laboratory, College of Veterinary Medicine, China Agricultural University, Beijing 100193, China"},{"name":"Key Laboratory of Animal Epidemiology of the Ministry of Agriculture, College of Veterinary Medicine, China Agricultural University, Beijing 100193, China"}],"role":[{"role":"author","vocabulary":"crossref"}]}],"member":"1968","published-online":{"date-parts":[[2021,7,26]]},"reference":[{"key":"ref_1","doi-asserted-by":"crossref","first-page":"944","DOI":"10.1016\/j.pt.2019.07.001","article-title":"Toxoplasma gondii","volume":"35","author":"Lourido","year":"2019","journal-title":"Trends Parasitol."},{"key":"ref_2","doi-asserted-by":"crossref","first-page":"1965","DOI":"10.1016\/S0140-6736(04)16412-X","article-title":"Toxoplasmosis","volume":"363","author":"Montoya","year":"2004","journal-title":"Lancet"},{"key":"ref_3","doi-asserted-by":"crossref","first-page":"542","DOI":"10.1016\/j.str.2015.01.010","article-title":"DNA-Damage-Inducible 1 Protein (Ddi1) Contains an Uncharacteristic Ubiquitin-like Domain that Binds Ubiquitin","volume":"23","author":"Urszula","year":"2015","journal-title":"Structure"},{"key":"ref_4","doi-asserted-by":"crossref","first-page":"818","DOI":"10.1126\/science.aax3769","article-title":"Cellular quality control by the ubiquitin-proteasome system and autophagy","volume":"366","author":"Pohl","year":"2019","journal-title":"Science"},{"key":"ref_5","doi-asserted-by":"crossref","first-page":"15790","DOI":"10.1073\/pnas.1521919112","article-title":"mTOR inhibition activates overall protein degradation by the ubiquitin proteasome system as well as by autophagy","volume":"112","author":"Zhao","year":"2015","journal-title":"Proc. Natl. Acad. Sci. USA"},{"key":"ref_6","doi-asserted-by":"crossref","first-page":"621","DOI":"10.1016\/j.chom.2015.10.014","article-title":"The Ubiquitin Proteome of Toxoplasma gondii Reveals Roles for Protein Ubiquitination in Cell-Cycle Transitions","volume":"18","author":"Yakubu","year":"2015","journal-title":"Cell Host Microbe"},{"key":"ref_7","doi-asserted-by":"crossref","first-page":"55","DOI":"10.1016\/S1471-4922(02)00064-8","article-title":"Characterization and role of protozoan parasite proteasomes","volume":"19","author":"Paugam","year":"2003","journal-title":"Trends Parasitol."},{"key":"ref_8","doi-asserted-by":"crossref","first-page":"3497","DOI":"10.1007\/s00018-016-2255-5","article-title":"The recognition of ubiquitinated proteins by the proteasome","volume":"73","author":"Grice","year":"2016","journal-title":"Cell Mol. Life Sci."},{"key":"ref_9","doi-asserted-by":"crossref","first-page":"1027","DOI":"10.1016\/j.jmb.2005.12.001","article-title":"UBL\/UBA ubiquitin receptor proteins bind a common tetraubiquitin chain","volume":"356","author":"Kang","year":"2006","journal-title":"J. Mol. Biol."},{"key":"ref_10","doi-asserted-by":"crossref","first-page":"291","DOI":"10.1146\/annurev-biochem-051810-094654","article-title":"Ubiquitin-binding proteins: Decoders of ubiquitin-mediated cellular functions","volume":"81","author":"Husnjak","year":"2012","journal-title":"Annu. Rev. Biochem."},{"key":"ref_11","doi-asserted-by":"crossref","first-page":"2819","DOI":"10.1007\/s00018-009-0048-9","article-title":"Ubiquitin-like and ubiquitin-associated domain proteins: Significance in proteasomal degradation","volume":"66","author":"Su","year":"2009","journal-title":"Cell Mol. Life Sci."},{"key":"ref_12","doi-asserted-by":"crossref","first-page":"5355","DOI":"10.1128\/MCB.25.13.5355-5362.2005","article-title":"The DNA damage-inducible UbL-UbA protein Ddi1 participates in Mec1-mediated degradation of Ho endonuclease","volume":"25","author":"Kaplun","year":"2005","journal-title":"Mol. Cell Biol."},{"key":"ref_13","doi-asserted-by":"crossref","first-page":"20390","DOI":"10.1074\/jbc.M110.112532","article-title":"The yeast E4 ubiquitin ligase Ufd2 interacts with the ubiquitin-like domains of Rad23 and Dsk2 via a novel and distinct ubiquitin-like binding domain","volume":"285","author":"Stingele","year":"2010","journal-title":"J. Biol. Chem."},{"key":"ref_14","doi-asserted-by":"crossref","first-page":"715","DOI":"10.1038\/35661","article-title":"Rad23 links DNA repair to the ubiquitin\/proteasome pathway","volume":"391","author":"Schauber","year":"1998","journal-title":"Nature"},{"key":"ref_15","doi-asserted-by":"crossref","first-page":"1331","DOI":"10.1083\/jcb.133.6.1331","article-title":"Yeast ubiquitin-like genes are involved in duplication of the microtubule organizing center","volume":"133","author":"Biggins","year":"1996","journal-title":"J. Cell Biol."},{"key":"ref_16","doi-asserted-by":"crossref","first-page":"139","DOI":"10.1016\/j.febslet.2010.11.026","article-title":"The retroviral proteinase active site and the N-terminus of Ddi1 are required for repression of protein secretion","volume":"585","author":"White","year":"2011","journal-title":"FEBS Lett."},{"key":"ref_17","doi-asserted-by":"crossref","first-page":"13711","DOI":"10.1096\/fj.202000759RR","article-title":"Toxoplasma gondii UBL-UBA shuttle proteins contribute to the degradation of ubiquitinylated proteins and are important for synchronous cell division and virulence","volume":"34","author":"Zhang","year":"2020","journal-title":"FASEB J."},{"key":"ref_18","doi-asserted-by":"crossref","first-page":"593","DOI":"10.1091\/mbc.01-06-0309","article-title":"Daughter Cell Assembly in the Protozoan Parasite Toxoplasma gondii","volume":"13","author":"Hu","year":"2002","journal-title":"Mol. Biol. Cell"},{"key":"ref_19","doi-asserted-by":"crossref","first-page":"1559","DOI":"10.1242\/jcs.021089","article-title":"Organellar dynamics during the cell cycle of Toxoplasma gondii","volume":"121","author":"Nishi","year":"2008","journal-title":"J. Cell Sci."},{"key":"ref_20","doi-asserted-by":"crossref","first-page":"1423","DOI":"10.1083\/jcb.151.7.1423","article-title":"The plastid of Toxoplasma gondii is divided by association with the centrosomes","volume":"151","author":"Striepen","year":"2000","journal-title":"J. Cell Biol."},{"key":"ref_21","doi-asserted-by":"crossref","first-page":"33","DOI":"10.1016\/j.devcel.2017.03.013","article-title":"Selective Autophagy of BES1 Mediated by DSK2 Balances Plant Growth and Survival","volume":"41","author":"Nolan","year":"2017","journal-title":"Dev. Cell"},{"key":"ref_22","doi-asserted-by":"crossref","unstructured":"Besteiro, S., Brooks, C.F., Striepen, B., and Dubremetz, J.F. (2011). Autophagy protein Atg3 is essential for maintaining mitochondrial integrity and for normal intracellular development of Toxoplasma gondii tachyzoites. PLoS Pathog., 7.","DOI":"10.1371\/journal.ppat.1002416"},{"key":"ref_23","doi-asserted-by":"crossref","first-page":"3625","DOI":"10.1091\/mbc.e07-05-0462","article-title":"Different domains of the UBL-UBA ubiquitin receptor, Ddi1\/Vsm1, are involved in its multiple cellular roles","volume":"19","author":"Gabriely","year":"2008","journal-title":"Mol. Biol. Cell"},{"key":"ref_24","doi-asserted-by":"crossref","first-page":"193","DOI":"10.1146\/annurev-biochem-061516-044908","article-title":"Proteasomal and Autophagic Degradation Systems","volume":"86","author":"Dikic","year":"2017","journal-title":"Annu. Rev. Biochem."},{"key":"ref_25","doi-asserted-by":"crossref","unstructured":"Liu, C., van Dyk, D., Li, Y., Andrews, B., and Rao, H. (2009). A genome-wide synthetic dosage lethality screen reveals multiple pathways that require the functioning of ubiquitin-binding proteins Rad23 and Dsk2. BMC Biol., 7.","DOI":"10.1186\/1741-7007-7-75"},{"key":"ref_26","doi-asserted-by":"crossref","first-page":"180","DOI":"10.3390\/cells3020180","article-title":"Yeast Irc22 Is a Novel Dsk2-Interacting Protein that Is Involved in Salt Tolerance","volume":"3","author":"Ishii","year":"2014","journal-title":"Cells"},{"key":"ref_27","doi-asserted-by":"crossref","first-page":"384","DOI":"10.1111\/1758-2229.12946","article-title":"Dsk2 involves in conidiation, multi-stress tolerance and thermal adaptation in Beauveria bassiana","volume":"13","author":"Wang","year":"2021","journal-title":"Environ. Microbiol. Rep."},{"key":"ref_28","doi-asserted-by":"crossref","first-page":"3001","DOI":"10.1007\/s00436-019-06449-w","article-title":"Triclosan inhibits the growth of Neospora caninum in vitro and in vivo","volume":"118","author":"Zhang","year":"2019","journal-title":"Parasitol. Res."},{"key":"ref_29","doi-asserted-by":"crossref","unstructured":"Wang, H., Lei, T., Liu, J., Li, M., Nan, H., and Liu, Q. (2014). A nuclear factor of high mobility group box protein in Toxoplasma gondii. PLoS ONE, 9.","DOI":"10.1371\/journal.pone.0111993"},{"key":"ref_30","doi-asserted-by":"crossref","first-page":"2082","DOI":"10.1093\/emboj\/cdg217","article-title":"Rapid invasion of host cells by Toxoplasma requires secretion of the MIC2-M2AP adhesive protein complex","volume":"22","author":"Huynh","year":"2003","journal-title":"EMBO J."},{"key":"ref_31","doi-asserted-by":"crossref","first-page":"1423","DOI":"10.1016\/j.cell.2016.08.019","article-title":"A Genome-wide CRISPR Screen in Toxoplasma Identifies Essential Apicomplexan Genes","volume":"166","author":"Sidik","year":"2016","journal-title":"Cell"}],"container-title":["International Journal of Molecular Sciences"],"original-title":[],"language":"en","link":[{"URL":"https:\/\/www.mdpi.com\/1422-0067\/22\/15\/7943\/pdf","content-type":"unspecified","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2025,10,11]],"date-time":"2025-10-11T06:34:55Z","timestamp":1760164495000},"score":1,"resource":{"primary":{"URL":"https:\/\/www.mdpi.com\/1422-0067\/22\/15\/7943"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[2021,7,26]]},"references-count":31,"journal-issue":{"issue":"15","published-online":{"date-parts":[[2021,8]]}},"alternative-id":["ijms22157943"],"URL":"https:\/\/doi.org\/10.3390\/ijms22157943","relation":{},"ISSN":["1422-0067"],"issn-type":[{"type":"electronic","value":"1422-0067"}],"subject":[],"published":{"date-parts":[[2021,7,26]]}}}