The histidine kinases of Candida albicans: regulation of cell wall mannan biosynthesis.
Previously, we have used both biochemical and immunological approaches to determine that the two-component, histidine kinase Chk1p regulates cell wall biosynthesis in Candida albicans. These data were obtained by comparing wild-type cells to a strain of C. albicans deleted in CHK1. The dysregulation of cell wall biosynthesis in the mutant reduces its adherence to human esophageal tissue and results in avirulence. In the current study, we used transmission immune electron microscopy (IEM) to visualize the cell surface of both wild-type (CAF2) and the chk1 mutant (CHK21). IEM was performed using two IgM monoclonal antibodies to either an acid-stable mannan epitope (Mab B6) or to an acid-labile mannan epitope (Mab B6.1). We observed that the cell surface of the CHK21 mutant was more reactive than wild-type cells with Mab B6, while the reactivity of Mab B6.1 was similar for both CAF2 and CHK21. These observations correlate with previous data on the Western blotting of mutant and wild-type cells using the same monoclonal antibodies, i.e., greater activity with Mab B6 than with Mab B6.1. In addition to CHK1, two other histidine kinases (SLN1 and NIK1) have been described in C. albicans. Mutants in both sln1Delta and nik1Delta were compared by Western blotting using Mab B6 and Mab B6.1. Reactivity of each mutant to Mab B6 was similar to that observed with the chk1 mutant; on the other hand, the mannoprotein profiles obtained with Mab B6.1 in all mutants were similar to wild-type cells. We also compared the expression of 29 genes involved in mannan synthesis by reverse transcription-polymerase chain reaction (RT-PCR) and found that expression of a subset of six genes (ALG2, ALG6, ALG8, MNT3, PMT6, KRT2) was upregulated in all histidine kinase mutants, while increased expression of ALG7 was only observed in the sln1 and nik1 mutants, MNN1 was upregulated in the chk1 and nik1 mutants, and MNN4 was upregulated in the nik1Delta. Our data indicate that each of the C. albicans HK proteins may regulate similar functions in cell wall biosynthesis. This activity could be achieved in either a common or parallel, redundant signal transduction pathway(s).
Kruppa
Michael
M
Department of Microbiology and Immunology, Georgetown University, Washington, DC, USA.
Jabra-Rizk
Mary Ann
MA
Meiller
Timothy F
TF
Calderone
Richard
R
D015153:Blotting, Western; D002176:Candida albicans / Q000201:enzymology* / Q000235:genetics; D002473:Cell Wall / Q000502:physiology*; D017353:Gene Deletion; D000071677:Histidine Kinase; D008351:Mannans / Q000096:biosynthesis*; D005810:Multigene Family; D011494:Protein Kinases / Q000235:genetics / Q000378:metabolism*; D015183:Restriction Mapping; D020133:Reverse Transcriptase Polymerase Chain Reaction
D008351:Mannans; D011494:Protein Kinases; D000071677:Histidine Kinase