Study On Ca Hsp90 And Ca Rlm1 Regulate Phenotypic Transition In Candida Albicans In Vitro

Objective:We aimed to study the Ca Hsp90 and Ca Rlml with CA ? regulate phenotypic transition in Candida albicans Methods:(1)lee's glucose and lee's GlcNAc containing different concentration of Dox(0,40,100,150?g/mL)was used for cell growth of Candida albicans.Cells were cultured at 25? and 37?.Filamentation in different treatments were observed.(2)Hsp90 was pharmacological inhibited with geldanamycin,in liquid medium with fermentable glycogen or non fermentable glycogen,the growth of the strain was observed and described growth curve of Candida albicans SC5314.(3)CDC19,TDH3 and PGK1 were cloned into the myc vector,and then was detected by Western blotting.(4)The over-expression of RLM1 with CA ?(11,21,or 30)were performed in GH1101(MTLa/?)and GH1322(MTLa/?).The phenotypes and white-opaque switching frequencies were observed.Results:(1)Treated on lee's glucose culture medium at 25? or 37?,low expression of Hsp90 induced the hyphae growth strongly;the ability to form hyphae decreased when increase of Hsp90 expression.Treated on lee's GlcNAc culture medium:? at 25? or 37?,increase of Hsp90 expression,GH1013 strain and HSP90 single knock-out strain had the hyphae growth ability radually decreased.? Treated at 25?,HSP90 conditional knockout strains hyphal growth ability weakened with Hsp90 expression increased.But,treated at 37?,there was no significant difference in the ability to form hyphae under different level of Hsp90 expression;and then HSP90 conditional knockout strains in GlcNAc culture medium was lower growth rate than glucose medium.(2)Candida albicans grew best in mannitol.Compromising Hsp90 function didn't affect utilization of glucose,fructose and mannitol.When GlcNAc was used as only glycogen,compromising Hsp90 function enhanced the cells apoptosis of Candida albicans.(3)The recombinant myc-CDC19,TDH3 and PGK1 vector were constructed successfully.Western blotting showed the expression of Myc-Cdc19 fusion protein in GH1013-tetO-HSP90/hsp90 cells.(4)There were no obvious phenotypic changes of RLM1 over-expression in GH1011 strains with different length of CA ?(CA ? 11,21 or 30).However,with the increase of the length of CA?,the switching frequency of white to opaque transition in GH1322 strain was increased.Conclusion:The phenotypic transition of Candida albicans is regulated by multiple genes.(1)The lower expression of Hsp90,the more strongerly induces yeast to filament transition.(2)The low expression of Hsp90 affects utilizing GlcNAc in Candida albicans.(3)The low expression of Hsp90 that GlcNAc accelerats the apoptosis of Candida albicans.(4)Hsp90 has no relationship with utilization of fermentable glycogen and non fermentable glycogen in Candida albicans.(5)The regulation ability of CA ?genotype of Candida albicans white to opaque morphological transformation is strain specificity.