NDLI: Isolation of Genes Mediating Resistance to Inhibitors of Nucleoside and Ergosterol Metabolism in Leishmania by Overexpression/Selection

Content Provider	Journal of Biological Chemistry (JBC)
Author	Cotrim, Paulo C. Garrity, Lynne K. Beverley, Stephen M.
Abstract	We tested a general method for the identification of drug resistance loci in the trypanosomatid protozoan parasiteLeishmania major. Genomic libraries in a multicopy episomal cosmid vector were transfected into susceptible parasites, and drug selections of these transfectant libraries yielded parasites bearing cosmids mediating resistance. Tests with two antifolates led to the recovery of cosmids encoding DHFR-TS or PTR1, two known resistance genes. Overexpression/selection using the toxic nucleoside tubercidin similarly yielded the TOR (toxic nucleoside resistance) locus, as well as a new locus (TUB2) conferring collateral hypersensitivity to allopurinol.Leishmania synthesize ergosterol rather than cholesterol, making this pathway attractive as a chemotherapeutic target. Overexpression/selection using the sterol synthesis inhibitors terbinafine (TBF, targeting squalene epoxidase) and itraconazole (ITZ, targeting lanosterol C14-demethylase) yielded nine new resistance loci. Several conferred resistance to both drugs; several were drug-specific, and two TBF-resistant cosmids induced hypersensitivity to ITZ. One TBF-resistant cosmid encoded squalene synthase (SQS1), which is located upstream of the sites of TBF and ITZ action in the ergosterol biosynthetic pathway. This suggests that resistance to “downstream” inhibitors can be mediated by increased expression of ergosterol biosynthetic intermediates. Our studies establish the feasibility of overexpression/selection in parasites and suggest that many Leishmania drug resistance loci are amenable to identification in this manner.
Related Links	http://www.jbc.org/content/274/53/37723.abstract
Ending Page	37730
Starting Page	37723
Page Count	8
File Format	HTM / HTML PDF
ISSN	00219258
Journal	Journal of Biological Chemistry (JBC)
Issue Number	53
Volume Number	274
DOI	10.1074/jbc.274.53.37723
e-ISSN	1083351X
Language	English
Publisher	American Society for Biochemistry and Molecular Biology
Publisher Date	1999-12-31
Access Restriction	Open
Subject Keyword	Methotrexate (MTX) Tubercidin (TUB) Itraconazole (ITZ) Terbinafine (TBF) Hygromycin B (HygB) Sodium dodecyl sulfate (SDS) Polymerase chain reaction (PCR) Kilobase pair (kb) NUCLEIC ACIDS, PROTEIN SYNTHESIS, AND MOLECULAR GENETICS
Content Type	Text
Resource Type	Article
Subject	Cell Biology Biochemistry Molecular Biology

Sl.	Authority	Responsibilities	Communication Details
1	Ministry of Education (GoI), Department of Higher Education	Sanctioning Authority	https://www.education.gov.in/ict-initiatives
2	Indian Institute of Technology Kharagpur	Host Institute of the Project: The host institute of the project is responsible for providing infrastructure support and hosting the project	https://www.iitkgp.ac.in
3	National Digital Library of India Office, Indian Institute of Technology Kharagpur	The administrative and infrastructural headquarters of the project	Dr. B. Sutradhar bsutra@ndl.gov.in
4	Project PI / Joint PI	Principal Investigator and Joint Principal Investigators of the project	Dr. B. Sutradhar bsutra@ndl.gov.in Prof. Saswat Chakrabarti will be added soon
5	Website/Portal (Helpdesk)	Queries regarding NDLI and its services	support@ndl.gov.in
6	Contents and Copyright Issues	Queries related to content curation and copyright issues	content@ndl.gov.in
7	National Digital Library of India Club (NDLI Club)	Queries related to NDLI Club formation, support, user awareness program, seminar/symposium, collaboration, social media, promotion, and outreach	clubsupport@ndl.gov.in
8	Digital Preservation Centre (DPC)	Assistance with digitizing and archiving copyright-free printed books	dpc@ndl.gov.in
9	IDR Setup or Support	Queries related to establishment and support of Institutional Digital Repository (IDR) and IDR workshops	idr@ndl.gov.in

Isolation of genes mediating resistance to inhibitors of nucleoside and ergosterol metabolism in leishmania by.

RNA Polymerase-specific Nucleosome Disruption by Transcription in Vivo

Inducible Expression of Suicide Genes in Leishmania donovani Amastigotes

Rescue of Embryonic Lethality in Reduced Folate Carrier-deficient Mice by Maternal Folic Acid Supplementation Reveals Early Neonatal Failure of Hematopoietic Organs

Identification of a Compound Origin of Replication at theHMR-E Locus in Saccharomyces cerevisiae

Editing of Cytochrome b mRNA inPhysarum Mitochondria

A New Family of Human Histone Deacetylases Related toSaccharomyces cerevisiae HDA1p

Dm1-MMP, a Matrix Metalloproteinase fromDrosophila with a Potential Role in Extracellular Matrix Remodeling during Neural Development

Cloning and Characterization of the Murine Ameloblastin Promoter

Isolation of Genes Mediating Resistance to Inhibitors of Nucleoside and Ergosterol Metabolism in Leishmania by Overexpression/Selection

Similar Documents

Isolation of genes mediating resistance to inhibitors of nucleoside and ergosterol metabolism in leishmania by.

RNA Polymerase-specific Nucleosome Disruption by Transcription in Vivo

Inducible Expression of Suicide Genes in Leishmania donovani Amastigotes

Rescue of Embryonic Lethality in Reduced Folate Carrier-deficient Mice by Maternal Folic Acid Supplementation Reveals Early Neonatal Failure of Hematopoietic Organs

Identification of a Compound Origin of Replication at theHMR-E Locus in Saccharomyces cerevisiae

Editing of Cytochrome b mRNA inPhysarum Mitochondria

A New Family of Human Histone Deacetylases Related toSaccharomyces cerevisiae HDA1p

Dm1-MMP, a Matrix Metalloproteinase fromDrosophila with a Potential Role in Extracellular Matrix Remodeling during Neural Development

Cloning and Characterization of the Murine Ameloblastin Promoter

Isolation of Genes Mediating Resistance to Inhibitors of Nucleoside and Ergosterol Metabolism in Leishmania by Overexpression/Selection