Characterization of a compensatory mutant of Leishmania major that lacks ether lipids but exhibits normal growth, and G418 and hygromycin resistance

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• 作者：Rachel Zufferey ; ^{zufferer@stjohns.edu} ; Stergios S. Bibis ; Tongtong Zhu ; Subbhalakshmi Dhalladoo
• 关键词：Leishmania major ; Ether glycerolipid ; Dihydroxyacetonephosphate acyltransferase ; Compensatory mutant ; Virulence ; Aminoglycoside resistance
• 刊名：Experimental Parasitology
• 出版年：2012
• 期刊代码：99_00144894
• 类别：imm
• 出版时间：March, 2012
• 卷：130
• 期：3
• 页码：200-204
• 文件大小：447 K

摘要

Ether glycerolipid biosynthesis in Leishmania major initiates with the acylation of dihydroxyacetonephosphate by the glycosomal dihydroxyacetonephosphate acyltransferase LmDAT. We previously reported that a null mutant of LmDAT 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, 螖lmdat/螖lmdat^rev. Similarly to the null mutant, the 螖lmdat/螖lmdat^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 antibiotics G418 and hygromycin was lower than that of the null mutant, wild type and complemented line.