Secreted lipases of Candida albicans: cloning, characterisation and expression analysis of a new gene family with at least ten members.
Extracellular lipolytic activity enabled the human pathogen Candida albicans to grow on lipids as the sole source of carbon. Nine new members of a lipase gene family (LIP2-LIP10) with high similarities to the recently cloned lipase gene LIP1 were cloned and characterised. The ORFs of all ten lipase genes are between 1281 and 1416 bp long and encode highly similar proteins with up to 80% identical amino acid sequences. Each deduced lipase sequence has conserved lipase motifs, four conserved cysteine residues, conserved putative N-glycosylation sites and similar hydrophobicity profiles. All LIP genes, except LIP7, also encode an N-terminal signal sequence. LIP3-LIP6 were expressed in all media and at all time points of growth tested as shown by Northern blot and RT-PCR analyses. LIP1, LIP3, LIP4, LIP5, LIP6 and LIP8 were expressed in medium with Tween 40 as a sole source of carbon. However, the same genes were also expressed in media without lipids. Two other genes, LIP2 and LIP9, were only expressed in media lacking lipids. Transcripts of most lipase genes were detected during the yeast-to-hyphal transition. Furthermore, LIP5, LIP6, LIP8 and LIP9 were found to be expressed during experimental infection of mice. These data indicate lipid-independent, highly flexible in vitro and in vivo expression of a large number of LIP genes, possibly reflecting broad lipolytic activity, which may contribute to the persistence and virulence of C. albicans in human tissue.
Hube
B
B
Robert Koch-Institut, Berlin, Germany. HubeB@rki.de
Stehr
F
F
Bossenz
M
M
Mazur
A
A
Kretschmar
M
M
Schäfer
W
W
D000595:Amino Acid Sequence; D002176:Candida albicans / Q000201:enzymology* / Q000235:genetics / Q000254:growth & development; D002177:Candidiasis / Q000382:microbiology; D003001:Cloning, Molecular; D003470:Culture Media; D005800:Genes, Fungal; D006801:Humans; D008049:Lipase / Q000737:chemistry / Q000235:genetics* / Q000378:metabolism*; D050356:Lipid Metabolism; D008969:Molecular Sequence Data; D005810:Multigene Family; D016366:Open Reading Frames; D020133:Reverse Transcriptase Polymerase Chain Reaction; D017422:Sequence Analysis, DNA
D003470:Culture Media; D008049:Lipase