TY - JOUR AU - Amoroso, G. AU - Morell-Avrahov, L. AU - Müller, D. AU - Klug, K. AU - Sültemeyer, D. TI - The gene NCE103 (YNL036w) from Saccharomyces cerevisiae encodes a functional carbonic anhydrase and its transcription is regulated by the concentration of inorganic carbon in the medium JO - Molecular microbiology VL - 56 SN - 0950-382X CY - Oxford [u.a.] PB - Wiley-Blackwell M1 - PreJuSER-46012 SP - 549 - 558 PY - 2005 N1 - Record converted from VDB: 12.11.2012 AB - Carbonic anhydrase (CA) catalyses the rapid interconversion between CO(2) and HCO(3) (-). Despite its wide distribution among living organisms, the presence of CA in fungi has been controversially discussed. Using mass spectrometric analysis of (18)O exchange from doubly labelled CO(2), we were able to measure CA activity in intact cells of Saccharomyces cerevisiae. Intracellular CA activity was lacking in the Deltance103 mutant, indicating that NCE103 encodes a functional CA. This was proven by overexpressing and purification of the NCE103 gene product showing a specific activity of around 6900 units per mg protein. Interestingly, the in vivo CA activity was 10-20 times higher in cells grown on low inorganic carbon (Ci; air containing 0.035% CO(2)) than in high-Ci cells (grown on 5% CO(2)). The enhanced CA activity of low-Ci cells was inducible after transferring high-Ci cells to air. Northern blot analysis revealed that that expression of NCE103 is transcriptionally regulated by low Ci which was also demonstrated by fusing the NCE103 promoter to beta-galactosidase as a reporter gene. Inactivation of NCE103 results in a high CO(2) requiring mutant indicating that a functional CA is an important prerequisite for S. cerevisiae to grow under low-Ci conditions. KW - Carbon: metabolism KW - Carbon: physiology KW - Carbonic Anhydrases: chemistry KW - Carbonic Anhydrases: classification KW - Carbonic Anhydrases: genetics KW - Carbonic Anhydrases: metabolism KW - Culture Media KW - Gene Expression Regulation, Fungal KW - Genes, Fungal KW - Saccharomyces cerevisiae: enzymology KW - Saccharomyces cerevisiae: genetics KW - Saccharomyces cerevisiae: metabolism KW - Transcription, Genetic: physiology KW - Culture Media (NLM Chemicals) KW - Carbon (NLM Chemicals) KW - Carbonic Anhydrases (NLM Chemicals) KW - J (WoSType) LB - PUB:(DE-HGF)16 C6 - pmid:15813743 UR - <Go to ISI:>//WOS:000227897400020 DO - DOI:10.1111/j.1365-2958.2005.04560.x UR - https://juser.fz-juelich.de/record/46012 ER -