The Candida albicans histidine kinase Chk1p: signaling and cell wall mannan.
Several published functions associated with the CHK1 histidine kinase of Candida albicans resemble those of the MAPK Cek1p and its cognate receptor Sho1p (SSU81). To explore this further, we have compared mutants lacking the proteins mentioned above and have constructed a double sho1/chk1Delta null mutant to determine relationships among these proteins. We observed that the sensitivity to Congo red (CR), calcofluor white (CW), as well as clumping of cells, was slightly increased in the double mutant compared to the single chk1Delta or sho1Delta mutants. However, Cek1p phosphorylation via Sho1p, which occurs during log phase growth in the presence or absence of CR in Wt cells, does not require Chk1p. These data suggest that Chk1p and Sho1p are components of parallel but independent signal pathways. In addition, bulk mannan of strains was analyzed by GLC/MS and GPC MALLS and NMR. Compared to Wt and a CHK1 gene-reconstituted strain (CHK23) that contained high, intermediate and low Mw mannan species, we found that the mannan of strains CHK21 (chk1Delta null), the cek1Delta null, and the double mutant consisted only of low Mw mannan. The sho1Delta null mutant only demonstrated a reduced intermediate type of mannan. Alcian blue binding was lower in cek1Delta, chk1Delta, and the double sho1/chk1Delta null mutant lacking high and intermediate Mw mannan than in the sho1Delta null which had a partial loss of intermediate Mw mannan only. We conclude that the Chk1p HK is part of a functionally similar but parallel pathway to the Sho1p-Cek1p pathway that confers resistance to the cell wall inhibitors CR and CW. However, a functional relationship in mannan biosynthesis of Chk1p and Cek1p exists that only partially requires Sho1p.
Li
Dongmei
D
Department of Microbiology and Immunology, Georgetown University Medical Center, Washington, DC 20057, USA.
Williams
David
D
Lowman
Douglas
D
Monteiro
Mario A
MA
Tan
Xuan
X
Kruppa
Michael
M
Fonzi
William
W
Roman
Elvira
E
Pla
Jesus
J
Calderone
Richard
R
D000935:Antifungal Agents / Q000494:pharmacology; D001557:Benzenesulfonates / Q000494:pharmacology; D002176:Candida albicans / Q000187:drug effects / Q000235:genetics / Q000502:physiology*; D002473:Cell Wall / Q000737:chemistry*; D000071877:Checkpoint Kinase 1; D003224:Congo Red / Q000494:pharmacology; D008401:Gas Chromatography-Mass Spectrometry; D017353:Gene Deletion; D015966:Gene Expression Regulation, Fungal; D005800:Genes, Fungal; D000071677:Histidine Kinase; D009682:Magnetic Resonance Spectroscopy; D008351:Mannans / Q000378:metabolism*; D011494:Protein Kinases / Q000235:genetics / Q000502:physiology*; D015398:Signal Transduction
D000935:Antifungal Agents; D001557:Benzenesulfonates; D008351:Mannans; D003224:Congo Red; C007061:C.I. Fluorescent Brightening Agent 28; D011494:Protein Kinases; D000071877:Checkpoint Kinase 1; D000071677:Histidine Kinase