Abstract
The upstream region of the pcbAB gene from Penicillium chrysogenum was screened for protein-binding sites using an electromobility shift assay. A specific protein/DNA interaction was detected within a fragment covering the region-387 to-242 relative to the pcbAB translational start codon. The appearance of this protein and pcbAB mRNA in culture extracts occurred at the same time point in fermentations, suggesting that the protein might be a transcription activator. The putative upstream activating sequence was located more precisely using cross-competition assays. These indicated the involvement of the 7-bp motif TGCCAAG in the binding of the protein.
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References
Bailey C, Arst HN Jr (1975) Carbon catabolite repression in Aspergillus nidulans. Eur J Biochem 51:574–577
Chu YW, Vichitsoonthonkul T, Keshavaz T, Renno D, Saunders G (1995) Molecular genetics as a tool to study the physiology of Penicillium chrysogenum grown in fermenters. Symposium of the 1995 IChemE research event/first European conference 2:1058–1060
Dhawale SS, Lane AC (1993) Compilation of sequence-specific DNA-binding proteins implicated in transcriptional control in fungi. Nucleic Acids Res 21:5537–5546
Díez B, Gutiérrez S, Barredo JL, van Solingen P, van der Voort LHM, Martín JF (1990) The cluster of penicillin biosynthetic genes. J Biol Chem 265:27:16358–16365
Espeso EA, Peñalva MA (1992) Carbon catabolite repression can account for the temporal pattern of expression of a penicillin biosynthetic gene in Aspergillus nidulans. Mol Microbiol 6:11: 1457–1465
Espeso EA, Tilburn J, Arst HN Jr, Peñalva MA (1993) pH regulation is a major determinant in expression of a fungal penicillin biosynthetic gene. EMBO J 12:10:3947–3956
Haas H, Redl B, Friedlin E, Stöffler G (1992) Isolation and analysis of the Penicillium chrysogenum phoA gene encoding a secreted phosphate-repressible acid phosphatase. Gene 113:129–133
Haas H, Friedlin E, Stöffler G, Redl B (1993) Cloning and structural organisation of a xylanase-encoding gene from Penicillium chrysogenum. Gene 126:237–242
Hersbach G, Van der Beek CP, Van Dijck PWM (1984) The penicillins: properties, biosynthesis, and fermentation. In: Vandamme EJ (ed) Biotechnology of industrial antibiotics. Marcel Dekker, New York, pp 45–140
Lein J (1986) The Panlabs strain improvement program. In: Vanek Z, Hostalek Z (eds) Overproduction of microbial metabolites. Butterworths, Boston, pp 105–139
MacCabe AP, Riach MBR, Unkles SE, Kinghorn JR (1990) The Aspergillus nidulans npeA locus consists of three contiguous genes required for penicillin biosynthesis. EMBO J 9:279–287
MacCabe AP, van Liempt H, Palissa H, Unkles SE, Riach MBR, Pfeifer E, von Döhren H, Kinghorn JR (1991) δ-(L-α-aminoadipolyl)-L-cysteinyl-D-valine synthetase from Aspergillus nidulans-molecular characterisation of the acvA gene encoding the first enzyme of the penicillin biosynthetic pathway. J Biol Chem 266:12646–12654
MacKenzie DA, Jeenes DJ, Belshaw NJ, Archer DB (1993) Regulation of secreted protein production by filamentous fungi: recent development and perspectives. J Gen Microbiol 139:2295–2307
Montenegro E, Barredo JL, Gutiérrez S, Díez B, Alvarez E, Martín JF (1990) Cloning, characterisation of the acyl-CoA:6-aminopenicillanic acid acyltransferase gene of Aspergillus nidulans and linkage to the isopenicillin N synthase gene. Mol Gen Genet 221:322–330
Pérez-Esteban B, Orejas M, Gómez-Pardo, Peñalva MA (1993) Molecular characterisation of a fungal secondary metabolism promoter: transcription of Aspergillus nidulans isopenicillin N synthetase gene is modulated by upstream negative elements. Mol Microbiol 9:4:881–895
Raper KB, Alexender DF (1945) Penicillin V: mycological aspects of penicillin production. J Elisha Mitchell Sci Soc 61:74–113
Renno DV, Saunders G, Bull AT, Holt G (1992) Transcript analysis of penicillin genes from Penicillium chrysogenum. Curr Genet 21:49–54
Shah AJ, Tilburn J, Adlard MW, Arst HN Jr (1993) pH regulation of penicillin production in Aspergillus nidulans. FEMS Microbiol Lett 77:209–212
Smith DJ, Earl AJ, Turner G (1990) The multifunctional peptide synthetase performing the first step of penicillin biosynthesis in Penicillium chrysogenum is a 421 073-dalton protein similar to Bacillus brevis antibiotic synthetases. EMBO J 9:2743–2750
Tilburn JT, Sarker S, Widdick DA, Espeso EA, Orejas M, Mungroo J, Peñalva MA, Arst HN Jr (1995) EMBO J (in press). The Aspergillus PacC zinc finger transcription factor mediates regulation of both acid and alkaline expressed genes by ambient pH. (1995) EMBO J 14, (4), 779–790
Turner G (1991) Cluster of genes involved in penicillin and cephalosporin biosynthesis. In: Kleinkauf H, von Dôhren H (eds) 50 years of penicillin application—history and trends. PUBLIC Ltd, Czech Republic, pp 115–124
Vichitsoonthonkul T, Chu YW, Sodhi H, Saunders G (1994) β-lactam antibiotics produced by genetically engineered filamentous fungi. In: Murooka Y, Imanaka T (eds) Recombinant microbes for industrial and agricultural applications. Marcel Dekker, New York, pp 119–135
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Chu, YW., Renno, D. & Saunders, G. Detection of a protein which binds specifically to the upstream region of the pcbAB gene in Penicillium chrysogenum. Curr Genet 28, 184–189 (1995). https://doi.org/10.1007/BF00315786
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DOI: https://doi.org/10.1007/BF00315786