Defect in dolichol-dependent glycosylation increases sensitivity of Saccharomyces cerevisiae towards anti-fungal drugs.
Two temperature-sensitive Saccharomyces cerevisiae mutants, sec59-1 and dpm1-6, impaired, respectively, in dolichol kinase (Sec59p) and dolichyl phosphate mannose (DolPMan) synthase (Dpm1p), have an aberrant cell wall structure and composition. We tested their sensitivity to four classes of antifungal drugs used in clinical practice: 5-fluorocytosine, amphotericin B, caspofungin and itraconasole. The strains were resistant to itraconazole and sensitive to the other drugs used. The minimal inhibitory concentration (MIC) of caspofungin and amphotericin B was two-fold lower for sec59-1 and dpm1-6 than for the respective wild-type strains. The sensitivity of both mutants could be brought back to the wild-type level by a multicopy suppressor of the thermosensitive phenotype, the RER2 gene, encoding cis-prenyltransferase involved in dolichol biosynthesis. Biochemical analysis revealed slight changes of the cell wall composition, different in the mutants as compared to the wild-type in response to the drugs. Our data strongly support a relationship between dolichol phosphate level, protein glycosylation and antifungal sensitivity.
Juchimiuk
Mateusz
M
Laboratory of Fungal Glycobiology, Institute of Biochemistry and Biophysics, Polish Academy of Sciences, Pawinskiego 5a, 02 106 Warsaw, Poland.
Pasikowska
Monika
M
Zatorska
Ewa
E
Laudy
Agnieszka E
AE
Smoleńska-Sym
Gabriela
G
Palamarczyk
Grazyna
G
D000935:Antifungal Agents / Q000494:pharmacology*; D002473:Cell Wall / Q000737:chemistry / Q000648:ultrastructure; D004122:Dimethylallyltranstransferase / Q000096:biosynthesis / Q000235:genetics; D004286:Dolichols / Q000378:metabolism*; D015870:Gene Expression; D006023:Glycoproteins / Q000378:metabolism*; D006031:Glycosylation; D006358:Hot Temperature; D008364:Mannosyltransferases / Q000172:deficiency*; D008826:Microbial Sensitivity Tests; D009154:Mutation; D017853:Phosphotransferases (Alcohol Group Acceptor) / Q000172:deficiency*; D012441:Saccharomyces cerevisiae / Q000187:drug effects* / Q000235:genetics / Q000378:metabolism*; D029701:Saccharomyces cerevisiae Proteins / Q000096:biosynthesis / Q000235:genetics
D000935:Antifungal Agents; D004286:Dolichols; D006023:Glycoproteins; D029701:Saccharomyces cerevisiae Proteins; D008364:Mannosyltransferases; C022755:dolichyl-phosphate beta-D-mannosyltransferase; D004122:Dimethylallyltranstransferase; C485360:RER2 protein, S cerevisiae; D017853:Phosphotransferases (Alcohol Group Acceptor); C020087:dolichol kinase