Abstract
The biosynthesis of penicillin, a secondary metabolite produced by Penicillium chrysogenum, is subject to sophisticated genetic and metabolic regulation. The structural genes, pcbC and pcbAB, which encode two of the penicillin biosynthetic enzymes are separated by a 1.16-kb intergenic region and transcribed divergently from one another. To identify and characterize nuclear proteins which interact with the pcbAB-pcbC intergenic promoter region, crude and partially purified nuclear extracts were used in mobility shift and DNA footprinting assays. Multiple DNA-binding proteins appear to bind to different regions of this DNA segment. An abundant nuclear protein, nuclear factor A (NF-A), binds at a single site in the intergenic promoter region and recognizes an 8-bp sequence, GCCAAGCC. Penicillin production is sensitive to nitrogen catabolite repression. The global-acting nitrogen regulatory protein NIT2 of Neurospora crassa binds strongly to the intergenic promoter region of the pcbAB and pcbC genes at a single site that contains two closely spaced GATA sequences.
This is a preview of subscription content, log in via an institution to check access.
Similar content being viewed by others
References
Barredo JL, van Solingen P, Diez B, Alvarez E, Cantoral JM, Kattevilder A, Smaal DB, Groenen MAM, Venstra AE, Martin JF (1989a) Gene 83:291–300
Barredo J, Cantoral JM, Alvarez E, Diez B, Martin JF (1989b) Mol Gen Genet 216:91–98
Brakhage AA, Browne P, Turner G (1992) J Bacteriol 174:3789–3799
Brunelle A, Schkeif RF (1987) Proc Natl Acad Sci USA 84:6773–6676
Car LG, Skatrud PL, Scheetzii ME, Queener SW, Ingolia TD (1986) Gene 48:257–266
Demain AL (1989) In: Hershberger C, Queener SW, Hegeman G (eds) Genetics and molecular biology of industrial microorganisms. Am Soc Microbiol, Washington DC, pp 1–11
Feng B, Xiao X, Marzluf GA (1993) Nucleic Acids Res 21:3989
Fu YH, Marzluf GA (1990a) Proc Natl Acad Sci USA 87:5331–5335
Fu YH, Marzluf GA (1990b) J Biol Chem 265:11 942–11 947
Gomez-Pardo E, Penalva MA (1990) J Biotechnol 16:109–115
Hautala JA, Conner BH, Jacobsen JW, Patel GL, Giles NH (1977) J Bacteriol 130:704–713
Jose ML, Revilla G, Villanueva JR, Martin JF (1979) J Gen Microbiol 115:207–211
Kadonaga JT, Tjian R (1986) Proc Natl Acad Sci USA 83:5889–5893
Kolar M, Holzmenn K, Weber G, Leitner E, Schwab H (1991) J Biotechnol 17:67–80
Kulmburg P, Mathieu M, Dower C, Kelly J, Felenbok B (1993) Mol Microbiol 7:847–857
MacCabe AP, Riach MB, Unkles SE, Kinghorn JR (1990) EMBO J 9:279–287
MacCabe AP, van Liempt H, Palissa H, Unkles SE, Riach MBR, Pfeifer E, von Dohren H, Kinghorn JR (1991) Delta-(L-alpha-aminoadipyl)-L-cysteinyl-D-valine synthetase from Aspergillus nidulans: molecular characterization of the acvA gene encoding the first enzyme of the penicillin biosynthetic pathway. J Biol Chem 266:12 646–12 654
Martin JF, Lopez-Nieto MJ, Castro JM, et al. (1986). Enzymes involved in beta-lactam biosynthesis controlled by carbon and nitrogen regulation. In: Kleinkauf I, von Doren H, Dornauer H, Nesemann G (eds) Regulation of secondary metabolite formation. VCH, Weinheim, pp 41–75
Matthews KS (1992) DNA looping. Microbiol Rev 56:123–136
Nehlin JO, Carlberg M, Ronne H (1991) Control of yeast GAL genes by MIG1 repressor: a transcriptional cascade in the glucose response. EMBO J 10:3373–3377
Perez-Esteban B, Orejas M, Gomez-Pardo E, Penalva MA (1993) Molecular characterization of a fungal secondary metabolism promoter: transcription of the Aspergillus nidulans isopenicillin N synthetase gene is modulated by upstream negative elements. Mol Microbiol 9:881–895
Renno DV, Saunders G, Bull AT, Holt G (1992) Transcript analysis of penicillin genes from Penicillium chrysogenum. Curr Genet 21:49–54
Revilla G, Ramos FR, Lopez-Nieto MJ, Martin JF (1986) J Antibiot 37:781–789
Sambrook J, Fritsch EF, Maniatis T (1989) Molecular cloning: a laboratory manual. Cold Spring Harbor Laboratory, Cold Spring Harbor, New York.
Sanchez S, Flores ME, Demain AL (1988) In: Sanchez-Esquivel (ed) Nitrogen source control of microbial processes. CRC Press, Boca Raton, Florida, pp 122–136
Smith DJ, Burnham MK, Bull JH, Hodgson JE, Ward JM, Browne P, Brown J, Barton B, Earl AJ, Turner G (1990) EMBO J 9: 741–747
Woodruff HB (1980) Science 208:1225–1229
Author information
Authors and Affiliations
Additional information
Communicated by K. Esser
Rights and permissions
About this article
Cite this article
Feng, B., Friedlin, E. & Marzluf, G.A. Nuclear DNA-binding proteins which recognize the intergenic control region of penicillin biosynthetic genes. Curr Genet 27, 351–358 (1995). https://doi.org/10.1007/BF00352104
Received:
Accepted:
Issue Date:
DOI: https://doi.org/10.1007/BF00352104