Modulation of gene expression in Leishmania drug resistant mutants as determined by targeted DNA microarrays

被引：98

作者：

Guimond, C

Trudel, N

Brochu, C

Marquis, N

El Fadili, A

Peytavi, R

Briand, G

Richard, D

Messier, N

Papadopoulou, B

Corbeil, J

Bergeron, MG

Légaré, D

Ouellette, M

机构：

[1] CHU Laval, Ctr Rech, Ctr Rech Infectiol, Quebec City, PQ G1V 4G2, Canada

[2] Univ Laval, Fac Med, Div Microbiol, Quebec City, PQ G1K 7P4, Canada

[3] Univ Calif San Diego, Sch Med, La Jolla, CA 92093 USA

来源：

NUCLEIC ACIDS RESEARCH | 2003年 / 31卷 / 20期

基金：

英国惠康基金; 加拿大健康研究院;

关键词：

D O I：

10.1093/nar/gkg806

中图分类号：

Q5 [生物化学]; Q7 [分子生物学];

学科分类号：

071010 ; 081704 ;

摘要：

In the protozoan parasite Leishmania, drug resistance can be a complex phenomenon. Several metabolic pathways and membrane transporters are implicated in the resistance phenotype. To monitor the expression of these genes, we generated custom DNA microarrays with PCR fragments corresponding to 44 genes involved with drug resistance. Transcript profiling of arsenite and antimony resistant mutants with these arrays pinpointed a number of genes overexpressed in mutants, including the ABC transporter PGPA, the glutathione biosynthesis genes gamma-glutamylcysteine synthetase (GSH1) and the glutathione synthetase (GSH2). Competitive hybridisations with total RNA derived from sensitive and methotrexate resistant cells revealed the overexpression of genes coding for dihydrofolate reductase (DHFR-TS), pteridine reductase (PTR1) and S-adenosylmethionine synthase (MAT2) and a down regulation of one gene of the folate transporter (FT) family. By labelling the DNA of sensitive and resistant parasites we could also detect several gene amplification events using DNA microarrays including the amplification of the S-adenosyl homocysteine hydrolase gene (SAHH). Alteration in gene expression detected by microarrays was validated by northern blot analysis, while Southern blots indicated that most genes overexpressed were also amplified, although other mechanisms were also present. The microarrays were useful in the study of resistant parasites to pinpoint several genes linked to drug resistance.

引用

页码：5886 / 5896

页数：11

共 56 条

[1]

Ben Mamoun C, 2001, MOL MICROBIOL, V39, P973, DOI 10.1046/j.1365-2958.2001.02289.x

[2] GENE AMPLIFICATION IN LEISHMANIA [J].

BEVERLEY, SM .

ANNUAL REVIEW OF MICROBIOLOGY, 1991, 45 :417-444

[3] Putting the Leishmania genome to work:: functional genomics by transposon trapping and expression profiling [J].

Beverley, SM ;

Akopyants, NS ;

Goyard, S ;

Matlib, RS ;

Gordon, JL ;

Brownstein, BH ;

Stormo, GD ;

Bukanova, EN ;

Hott, CT ;

Li, FG ;

MacMillan, S ;

Muo, JN ;

Schwertman, LA ;

Smeds, MR ;

Wang, YJ .

PHILOSOPHICAL TRANSACTIONS OF THE ROYAL SOCIETY OF LONDON SERIES B-BIOLOGICAL SCIENCES, 2002, 357 (1417) :47-53

[4]

BORST P, 1995, ANNU REV MICROBIOL, V49, P427, DOI 10.1146/annurev.mi.49.100195.002235

[5] Expression profiling of the schizont and trophozoite stages of Plasmodium falciparum with a long-oligonucleotide microarray -: art. no. R9 [J].

Bozdech, Z ;

Zhu, JC ;

Joachimiak, MP ;

Cohen, FE ;

Pulliam, B ;

DeRisi, JL .

GENOME BIOLOGY, 2003, 4 (02)

[6]

BROCHU C, 2003, IN PRESS ANTIMICROB

[7]

CALLAHAN HL, 1992, J BIOL CHEM, V267, P24165

[8] Treatment-specific changes in gene expression discriminate in vivo drug response in human leukemia cells [J].

Cheok, MH ;

Yang, WL ;

Pui, CH ;

Downing, JR ;

Cheng, C ;

Naeve, CW ;

Relling, MV ;

Evans, WE .

NATURE GENETICS, 2003, 34 (01) :85-90

[9] OVERPRODUCTION OF A BIFUNCTIONAL THYMIDYLATE SYNTHETASE-DIHYDROFOLATE REDUCTASE AND DNA AMPLIFICATION IN METHOTREXATE-RESISTANT LEISHMANIA-TROPICA [J].

CODERRE, JA ;

BEVERLEY, SM ;

SCHIMKE, RT ;

SANTI, DV .

PROCEEDINGS OF THE NATIONAL ACADEMY OF SCIENCES OF THE UNITED STATES OF AMERICA-BIOLOGICAL SCIENCES, 1983, 80 (08) :2132-2136

[10] Population genomics of drug resistance in Candida albicans [J].

Cowen, LE ;

Nantel, A ;

Whiteway, MS ;

Thomas, DY ;

Tessier, DC ;

Kohn, LM ;

Anderson, JB .

PROCEEDINGS OF THE NATIONAL ACADEMY OF SCIENCES OF THE UNITED STATES OF AMERICA, 2002, 99 (14) :9284-9289

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