دورية أكاديمية
أعرض في EDS

Nucleoside Transport and Nucleobase Uptake Null Mutants in Leishmania mexicana for the Routine Expression and Characterization of Purine and Pyrimidine Transporters.

التفاصيل البيبلوغرافية
العنوان: Nucleoside Transport and Nucleobase Uptake Null Mutants in Leishmania mexicana for the Routine Expression and Characterization of Purine and Pyrimidine Transporters.
المؤلفون: Aldfer MM; Institute of Infection, Immunity and Inflammation, College of Medical, Veterinary and Life Sciences, University of Glasgow, Glasgow G12 8QQ, UK., AlSiari TA; Institute of Infection, Immunity and Inflammation, College of Medical, Veterinary and Life Sciences, University of Glasgow, Glasgow G12 8QQ, UK., Elati HAA; Institute of Infection, Immunity and Inflammation, College of Medical, Veterinary and Life Sciences, University of Glasgow, Glasgow G12 8QQ, UK., Natto MJ; Institute of Infection, Immunity and Inflammation, College of Medical, Veterinary and Life Sciences, University of Glasgow, Glasgow G12 8QQ, UK., Alfayez IA; Institute of Infection, Immunity and Inflammation, College of Medical, Veterinary and Life Sciences, University of Glasgow, Glasgow G12 8QQ, UK., Campagnaro GD; Institute of Infection, Immunity and Inflammation, College of Medical, Veterinary and Life Sciences, University of Glasgow, Glasgow G12 8QQ, UK., Sani B; Institute of Infection, Immunity and Inflammation, College of Medical, Veterinary and Life Sciences, University of Glasgow, Glasgow G12 8QQ, UK., Burchmore RJS; Institute of Infection, Immunity and Inflammation, College of Medical, Veterinary and Life Sciences, University of Glasgow, Glasgow G12 8QQ, UK., Diallinas G; Department of Biology, National and Kapodistrian University of Athens, Panepistimioupolis, 15784 Athens, Greece.; Institute of Molecular Biology and Biotechnology, Foundation for Research and Technology, 70013 Heraklion, Greece., De Koning HP; Institute of Infection, Immunity and Inflammation, College of Medical, Veterinary and Life Sciences, University of Glasgow, Glasgow G12 8QQ, UK.
المصدر: International journal of molecular sciences [Int J Mol Sci] 2022 Jul 23; Vol. 23 (15). Date of Electronic Publication: 2022 Jul 23.
نوع المنشور: Journal Article
اللغة: English
بيانات الدورية: Publisher: MDPI Country of Publication: Switzerland NLM ID: 101092791 Publication Model: Electronic Cited Medium: Internet ISSN: 1422-0067 (Electronic) Linking ISSN: 14220067 NLM ISO Abbreviation: Int J Mol Sci Subsets: MEDLINE
أسماء مطبوعة: Original Publication: Basel, Switzerland : MDPI, [2000-
مواضيع طبية MeSH: Leishmania mexicana*/genetics, Biological Transport ; Equilibrative Nucleoside Transport Proteins/metabolism ; Membrane Transport Proteins/genetics ; Membrane Transport Proteins/metabolism ; Nucleosides/metabolism ; Purines/metabolism ; Pyrimidines/metabolism ; Uracil/metabolism
مستخلص: The study of transporters is highly challenging, as they cannot be isolated or studied in suspension, requiring a cellular or vesicular system, and, when mediated by more than one carrier, difficult to interpret. Nucleoside analogues are important drug candidates, and all protozoan pathogens express multiple equilibrative nucleoside transporter (ENT) genes. We have therefore developed a system for the routine expression of nucleoside transporters, using CRISPR/cas9 to delete both copies of all three nucleoside transporters from Leishmania mexicana (ΔNT1.1/1.2/2 (SUPKO)). SUPKO grew at the same rate as the parental strain and displayed no apparent deficiencies, owing to the cells' ability to synthesize pyrimidines, and the expression of the LmexNT3 purine nucleobase transporter. Nucleoside transport was barely measurable in SUPKO, but reintroduction of L. mexicana NT1.1, NT1.2, and NT2 restored uptake. Thus, SUPKO provides an ideal null background for the expression and characterization of single ENT transporter genes in isolation. Similarly, an LmexNT3-KO strain provides a null background for transport of purine nucleobases and was used for the functional characterization of T. cruzi NB2, which was determined to be adenine-specific. A 5-fluorouracil-resistant strain (Lmex5FURes) displayed null transport for uracil and 5FU, and was used to express the Aspergillus nidulans uracil transporter FurD.
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معلومات مُعتمدة: 206385/2014-5 National Council for Scientific and Technological Development; PhD studentship (2) Saudi Arabian government; PhD studentship (2) Libyan government; Institutional Strategic Support Fund (ISSF) Award (ref 317019) United Kingdom WT_ Wellcome Trust
فهرسة مساهمة: Keywords: Aspergillus nidulans; FurD uracil transporter; Trichomonas vaginalis; Trypanosoma cruzi; equilibrative nucleoside transporter; expression system; nucleobase transport
المشرفين على المادة: 0 (Equilibrative Nucleoside Transport Proteins)
0 (Membrane Transport Proteins)
0 (Nucleosides)
0 (Purines)
0 (Pyrimidines)
56HH86ZVCT (Uracil)
تواريخ الأحداث: Date Created: 20220728 Date Completed: 20220729 Latest Revision: 20220731
رمز التحديث: 20221213
مُعرف محوري في PubMed: PMC9331716
DOI: 10.3390/ijms23158139
PMID: 35897714
قاعدة البيانات: MEDLINE
الوصف
تدمد:1422-0067
DOI:10.3390/ijms23158139