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In this study, we have characterized the biochemical and functional role of a novel ubiquitin-specific protease (USP) domain-containing protein of the human malaria parasite Plasmodium falciparum (PfUSP). We have shown that the PfUSP is an active deubiquitinase associated with parasite endoplasmic reticulum (ER). Selection linked integration (SLI) method for C-terminal tagging and GlmS-ribozyme mediated inducible knock-down (iKD) of PfUSP was utilized to assess its functional role. Inducible knockdown of PfUSP resulted in a remarkable reduction in parasite growth and multiplication; specifically, PfUSP-iKD disrupted ER morphology and development, blocked the development of healthy schizonts, and hindered proper merozoite development. PfUSP-iKD caused increased ubiquitylation of specific proteins, disrupted organelle homeostasis and reduced parasite survival. Since the mode of action of artemisinin and the artemisinin-resistance are shown to be associated with the proteasome machinery, we analyzed the effect of dihydroartemisinin (DHA) on PfUSP-iKD parasites. Importantly, the PfUSP-knocked-down parasite showed increased sensitivity to dihydroartemisinin (DHA), whereas no change in chloroquine sensitivity was observed, suggesting a role of PfUSP in combating artemisinin-induced cellular stress. Together, the results show that Plasmodium PfUSP is an essential protease for parasite survival, and its inhibition increases the efficacy of artemisinin-based drugs. Therefore, PfUSP can be targeted to develop novel scaffolds for developing new antimalarials to combat artemisinin resistance.<\/jats:p>","DOI":"10.1042\/bcj20220429","type":"journal-article","created":{"date-parts":[[2022,12,13]],"date-time":"2022-12-13T13:48:42Z","timestamp":1670939322000},"page":"25-39","update-policy":"https:\/\/doi.org\/10.1042\/crossmark_policy","source":"Crossref","is-referenced-by-count":7,"title":["A <i>Plasmodium falciparum<\/i> ubiquitin-specific protease (<i>Pf<\/i>USP) is essential for parasite survival and its disruption enhances artemisinin efficacy"],"prefix":"10.1042","volume":"480","author":[{"given":"Priya","family":"Arora","sequence":"first","affiliation":[{"name":"International Centre for Genetic Engineering and Biotechnology, New Delhi 110 067, India"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Monika","family":"Narwal","sequence":"additional","affiliation":[{"name":"International Centre for Genetic Engineering and Biotechnology, New Delhi 110 067, India"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Vandana","family":"Thakur","sequence":"additional","affiliation":[{"name":"International Centre for Genetic Engineering and Biotechnology, New Delhi 110 067, India"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Osama","family":"Mukhtar","sequence":"additional","affiliation":[{"name":"International Centre for Genetic Engineering and Biotechnology, New Delhi 110 067, India"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Pawan","family":"Malhotra","sequence":"additional","affiliation":[{"name":"International Centre for Genetic Engineering and Biotechnology, New Delhi 110 067, India"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-0239-1979","authenticated-orcid":false,"given":"Asif","family":"Mohmmed","sequence":"additional","affiliation":[{"name":"International Centre for Genetic Engineering and Biotechnology, New Delhi 110 067, India"}],"role":[{"role":"author","vocabulary":"crossref"}]}],"member":"288","published-online":{"date-parts":[[2023,1,6]]},"reference":[{"key":"2023010614401699500_BCJ-2022-0429C1","unstructured":"World Health Organization. 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