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Characterization of a compensatory mutant of Leishmania major that lacks ether lipids but exhibits normal growth and G418 resistance
Author(s) -
Zhu Tongtong,
Dhalladoo Subbhalaskshmi,
Bibis Sergio,
Zufferey Rachel
Publication year - 2012
Publication title -
the faseb journal
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.709
H-Index - 277
eISSN - 1530-6860
pISSN - 0892-6638
DOI - 10.1096/fasebj.26.1_supplement.623.1
Subject(s) - mutant , biology , lipophosphoglycan , complementation , microbiology and biotechnology , acyltransferase , mutation , null allele , biochemistry , gene , genetics , leishmania donovani , leishmaniasis , visceral leishmaniasis
Ether glycerolipid biosynthesis in Leishmania major initiates with the acylation of dihydroxyacetonephosphate by the glycosomal dihydroxyacetonephosphate acyltransferase Lm DAT. We previously reported that a null mutant of Lm DAT is severely affected in logarithmic growth, survival during stationary phase, and in virulence in mice. In addition, it lacks all ether glycerolipids, produces altered forms of the ether‐lipid based virulence factors lipophosphoglycan and increased levels of GPI‐anchored protein gp63. Here, we describe the characterization of a compensatory mutant of a null strain of LmDAT , Δlmdat/Δlmdat rev . Similarly to the null mutant, the Δlmdat/Δlmdat rev strain formed altered forms of lipophosphoglycan and increased levels of gp63, and was avirulent in mice infection. Further, dihydroxyacetonephosphate acyltransferase activity was absent in the revertant clone, indicating that a mutation in another acyltransferase gene did not confer dihydroxyacetonephosphate specificity. In contrast, the revertant grew normally but still exhibited poor survival during stationary phase. In addition, agarose gel analysis of its genomic DNA failed to detect any amplified DNA. Surprisingly, its sensitivity to aminoglycoside based antibiotic G418 was three times lower than that of the null mutant, while resistance to blasticidin and hygromycin was similar to that of the null mutant.