Summary
Some of the properties of the RNA polymerase purified from SPO1-infected Bacillus subtilis have been compared with the properties of RNA polymerase from uninfected cells (core+δ). The two enzymes synthetize RNA from nonoverlapping regions on SPO1 DNA, and they lead to the retention of different restriction fragments of SPO1 DNA on cellulosenitrate filters.
The action of the positively regulating product of gene 28 of SPO1 (gp 28) has been analyzed. The isolated gp 28 has been shown to be unable to increase the dissociation rate of core +δ from SPP1 DNA, while it efficiently blocks the initiation of RNA synthesis if it is added to preformed complexes between core+δ and SPP1 DNA in 0.2 M NaCl.
This is a preview of subscription content, log in via an institution to check access.
Similar content being viewed by others
References
Chaconas G, Van de Sande JH (1980) 5′-32P labeling of RNA and DNA restriction fragments. In: Grossman L, Moldave K (eds) Methods in enzymology, vol 65. Academic Press, New York, pp 75–88
Duffy JJ, Geiduschek EP (1975) RNA Polymerase from Phage SPO1 infected and uninfected B. subtilis. J Biol Chem 250:4530–4541
Duffy JJ, Geiduschek EP (1977) Purification of a positive regulatory subunit from phage SPO1 modified RNA polymerase. Nature 270:28–32
Gabain A von, Bujard H (1977) Interaction of E. coli RNA polymerase with promoters of coliphage T5. Mol Gen Genet 157:301–311
Gage PL, Geiduschek EP (1971) RNA synthesis during bacteriophage SPO1 Development: Six classes of SPO1 RNA. J Mol Biol 57:279–300
Giacomoni PU (1980) Purification and DNA-Binding Properties of RNA polymerase from B. subtilis. Eur J Biochem 106:579–591
Giacomoni PU (1981a) Protein-nucleic acid interaction. I.: An empirical approach to the filter retention assay. Biochem International 2:389–397
Giacomoni PU (1981b) Protein-nucleid acid interaction. 2: A rational approach to the filter retention assay. Biochem International 2:399–410
Giacomoni PU, Delain E, Le Pecq JB (1977) Electron microscopy analysis of the interaction between. E. coli DNA-dependent RNA polymerase and the replicative form of phage fd DNA. 2. Analysis of the dissociation kinetics. Eur J Biochem 78:215–220
Hinkle DC, Chamberlin MJ (1972) Studies of the binding of E. coli RNA polymerase to DNA. I. The role of sigma subunit in site selection. J Mol Biol 70:157–185
Hinkle DC, Mangel WF, Chamberlin MJ (1972) Studies of the binding of E. coli RNA polymerase to DNA. IV. The effect of rifampicin on binding and on RNA chain initiation. J Mol Biol 70:209–220
McClure WR, Cech CL (1978). On the mechanism of Rifampicin Inhibition of RNA Synthesis. J Biol Chem 253:8949–8956
Riva S, Polsinelli M, Falaschi A (1968) A new phage of B. subtilis with infectious DNA having separable strands. J Mol Biol 35:347–356
Seeburg PH, Nüsslein Ch, Schaller H (1977) Interaction of RNA polymerase with promoters from bacteriophage fd. Eur J Biochem 74:107–113
Southern EM (1975) Detection of specific sequences among DNA fragments separated by gel electrophoresis. J Mol Biol 98:503–517
Williamson VM, Doi RH (1979) Sigma factor is not released during transcription in B. subtilis. Mol Gen Genet 174:47–52
Author information
Authors and Affiliations
Additional information
Communicated by E. Bautz
Rights and permissions
About this article
Cite this article
Giacomoni, P.U. Effects of the positively regulating product of gene 28 of the B. subtilis phage SPO1 on in vitro transcription. Molec. Gen. Genet. 183, 422–427 (1981). https://doi.org/10.1007/BF00268760
Received:
Issue Date:
DOI: https://doi.org/10.1007/BF00268760