Identification and purification of CREB like protein in Candida albicans.
cAMP response element binding protein (CREB) belongs to ATF/CREB family of transcription factors, which are bonafide targets of cAMP-PKA signalling pathway in mammalian cells. CREB is known to regulate the genes involved in transcription, cell cycle, cell survival, neurotransmitter, growth factors and immune regulation. But there is no evidence of presence of ATF/CREB family members in Candida albicans. In the present study, CREB like transcription factor has been identified and purified in C. albicans. The putative CREB was observed to have different molecular mass (47 kDa) as compared to its mammalian counterpart (43 kDa). Both forms of CREB (CREB and phosphorylated CREB) were detected in C. albicans and phosphorylation of CREB was found to be a function of cAMP levels and protein kinase A activity within this organism. CREB protein was purified by sequence-specific CRE-DNA affinity chromatography. Purified CREB exhibited characteristic CRE binding activity as revealed by electrophoretic mobility shift assay and gave reactivity with CREB antibodies. CREB protein was phosphorylated by purified catalytic subunit of PKA under in vitro conditions. To the best of our knowledge, this study reports for the first time identification of CREB like protein as an important component of cAMP signalling pathway in C. albicans.
Singh
Ajay
A
Department of Biochemistry, Postgraduate Institute of Medical Education & Research, Chandigarh 160 012, India.
Dhillon
Navneet K
NK
Sharma
Sadhna
S
Khuller
Gopal K
GK
D002176:Candida albicans / Q000187:drug effects / Q000201:enzymology / Q000378:metabolism*; D020134:Catalytic Domain; D000242:Cyclic AMP / Q000494:pharmacology; D017362:Cyclic AMP Response Element-Binding Protein / Q000302:isolation & purification*; D017868:Cyclic AMP-Dependent Protein Kinases / Q000378:metabolism; D010750:Phosphoproteins / Q000378:metabolism; D010766:Phosphorylation / Q000187:drug effects
D017362:Cyclic AMP Response Element-Binding Protein; D010750:Phosphoproteins; D000242:Cyclic AMP; D017868:Cyclic AMP-Dependent Protein Kinases