PMID- 36625569 OWN - NLM STAT- MEDLINE DCOM- 20230222 LR - 20230711 IS - 1098-6596 (Electronic) IS - 0066-4804 (Print) IS - 0066-4804 (Linking) VI - 67 IP - 2 DP - 2023 Feb 16 TI - A Plasmodium falciparum RING Finger E3 Ubiquitin Ligase Modifies the Roles of PfMDR1 and PfCRT in Parasite Drug Responses. PG - e0082122 LID - 10.1128/aac.00821-22 [doi] LID - e00821-22 AB - Protein ubiquitination is an important posttranslational regulation mechanism that mediates Plasmodium development and modifies parasite responses to antimalarial drugs. Although mutations in several parasite ubiquitination enzymes have been linked to increased drug tolerance, the molecular mechanisms by which ubiquitination pathways mediate these parasite responses remain largely unknown. Here, we investigate the roles of a Plasmodium falciparum ring finger ubiquitin ligase (PfRFUL) in parasite development and in responses to antimalarial drugs. We engineered a transgenic parasite having the Pfrful gene tagged with an HA-2A-NeoR-glmS sequence to knockdown (KD) Pfrful expression using glucosamine (GlcN). A Western blot analysis of the proteins from GlcN-treated pSLI-HA-NeoR-glmS-tagged (PfRFULg) parasites, relative to their wild-type (Dd2) controls, showed changes in the ubiquitination of numerous proteins. PfRFUL KD rendered the parasites more sensitive to multiple antimalarial drugs, including mefloquine, piperaquine, amodiaquine, and dihydroartemisinin. PfRFUL KD also decreased the protein level of the P. falciparum multiple drug resistance 1 protein (PfMDR1) and altered the ratio of two bands of the P. falciparum chloroquine resistance transporter (PfCRT), suggesting contributions to the changed drug responses by the altered ubiquitination of these two molecules. The inhibition of proteasomal protein degradation by epoxomicin increased the PfRFUL level, suggesting the degradation of PfRFUL by the proteasome pathways, whereas the inhibition of E3 ubiquitin ligase activities by JNJ26854165 reduced the PfRFUL level. This study reveals the potential mechanisms of PfRFUL in modifying the expression of drug transporters and their roles in parasite drug responses. PfRFUL could be a potential target for antimalarial drug development. FAU - Singh, Brajesh K AU - Singh BK AD - Laboratory of Malaria and Vector Research, National Institute of Allergy and Infectious Diseases, National Institutes of Health, Rockville, Maryland, USA. FAU - Zhang, Cui AU - Zhang C AD - Laboratory of Malaria and Vector Research, National Institute of Allergy and Infectious Diseases, National Institutes of Health, Rockville, Maryland, USA. FAU - Wu, Jian AU - Wu J AD - Laboratory of Malaria and Vector Research, National Institute of Allergy and Infectious Diseases, National Institutes of Health, Rockville, Maryland, USA. FAU - Peng, Yu-Chih AU - Peng YC AD - Laboratory of Malaria and Vector Research, National Institute of Allergy and Infectious Diseases, National Institutes of Health, Rockville, Maryland, USA. FAU - He, Xiao AU - He X AD - Laboratory of Malaria and Vector Research, National Institute of Allergy and Infectious Diseases, National Institutes of Health, Rockville, Maryland, USA. FAU - Tumas, Keyla C AU - Tumas KC AD - Laboratory of Malaria and Vector Research, National Institute of Allergy and Infectious Diseases, National Institutes of Health, Rockville, Maryland, USA. FAU - Sa, Juliana M AU - Sa JM AD - Laboratory of Malaria and Vector Research, National Institute of Allergy and Infectious Diseases, National Institutes of Health, Rockville, Maryland, USA. FAU - Lane, Kristin D AU - Lane KD AD - Laboratory of Malaria and Vector Research, National Institute of Allergy and Infectious Diseases, National Institutes of Health, Rockville, Maryland, USA. FAU - Eastman, Richard T AU - Eastman RT AD - Laboratory of Malaria and Vector Research, National Institute of Allergy and Infectious Diseases, National Institutes of Health, Rockville, Maryland, USA. AD - Early Translation Branch, National Center for Advancing Translational Sciences (NCATS), National Institutes of Health, Rockville, Maryland, USA. FAU - Narum, David L AU - Narum DL AD - Laboratory of Malaria Immunology and Vaccinology, National Institute of Allergy and Infectious Diseases, National Institutes of Health, Bethesda, Maryland, USA. FAU - Wellems, Thomas E AU - Wellems TE AUID- ORCID: 0000-0003-3899-8454 AD - Laboratory of Malaria and Vector Research, National Institute of Allergy and Infectious Diseases, National Institutes of Health, Rockville, Maryland, USA. FAU - Su, Xin-Zhuan AU - Su XZ AUID- ORCID: 0000-0003-3246-3248 AD - Laboratory of Malaria and Vector Research, National Institute of Allergy and Infectious Diseases, National Institutes of Health, Rockville, Maryland, USA. LA - eng PT - Journal Article PT - Research Support, N.I.H., Intramural DEP - 20230110 PL - United States TA - Antimicrob Agents Chemother JT - Antimicrobial agents and chemotherapy JID - 0315061 RN - 0 (Antimalarials) RN - 886U3H6UFF (Chloroquine) RN - 0 (Membrane Transport Proteins) RN - 0 (Multidrug Resistance-Associated Proteins) RN - 0 (Protozoan Proteins) RN - EC 2.3.2.27 (Ubiquitin-Protein Ligases) SB - IM MH - Humans MH - *Antimalarials/pharmacology MH - Chloroquine/pharmacology MH - Drug Resistance/genetics MH - Malaria, Falciparum/drug therapy MH - Membrane Transport Proteins/genetics MH - Multidrug Resistance-Associated Proteins/genetics MH - *Plasmodium falciparum/drug effects/genetics MH - *Protozoan Proteins/genetics/metabolism MH - *Ubiquitin-Protein Ligases/genetics/metabolism PMC - PMC9933707 OTO - NOTNLM OT - drug responses OT - gene knockdown OT - malaria OT - protein ubiquitination OT - rodent COIS- The authors declare no conflict of interest. EDAT- 2023/01/11 06:00 MHDA- 2023/02/22 06:00 PMCR- 2023/07/10 CRDT- 2023/01/10 09:03 PHST- 2023/01/10 09:03 [entrez] PHST- 2023/02/22 06:00 [medline] PHST- 2023/01/11 06:00 [pubmed] PHST- 2023/07/10 00:00 [pmc-release] AID - 00821-22 [pii] AID - aac.00821-22 [pii] AID - 10.1128/aac.00821-22 [doi] PST - ppublish SO - Antimicrob Agents Chemother. 2023 Feb 16;67(2):e0082122. doi: 10.1128/aac.00821-22. Epub 2023 Jan 10.