Absence of functional RNA encoded by a silent chromosome in non-complementing diplods obtained from protoplast fusion in Bacillus subtilis
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The molecular basis for lack of phenotypic expression of one chromosome in Bacillus subtilis non-complementing diploid clones (Ncd cells) was investigated.
Correlations between chromosomal inactivation and absence of functional transcripts were determined with wild-type prophage ϕ105 or a thermoinducible mutant ϕ105 cts23, on either the expressed or the silent chromosome.
It appears that no significant amount of phage mRNA is detectable in Ncd cells carrying the prophage in the inactive chromosome. However, ϕ105 mRNA represents 0.23% of total cellular mRNA in an Ncd strain with the prophage in the expressed chromosome and 0.28% in the parental lysogenic strain.
The lack of an mRNA repressor of ϕ105 prophage from the silent chromosome was confirmed by the absence of repressor activity in Ncd clones with a temperature sensitive mutant ϕ105 located in the silent chromosome. After heat induction, no phage production was observed. As expected these clones do not exhibit ϕ105 immunity when superinfected with the same phage.
The combined data of the present and previous work suggest that control of phenotypic suppression of Ncd strains should, at the transcription level, involve a different DNA tertiary organisation in one of the two chromosomes.
KeywordsBacillus Subtilis Transcription Level Phenotypic Expression Protoplast Fusion Chromosomal Inactivation
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- Anagnostopoulos C, Spizizen J (1961) Requirement for transformation in B. subtilis. J Bacteriol 81:741–746Google Scholar
- Armentrout R, Rutberg L (1970) Mapping of prophage and mature deoxyribonucleic acid from temperate Bacillus bacteriophage ϕ105 by marker rescue. J Virol 6:760–767Google Scholar
- Bohin JP, Ben Khalifa K, Guillén N, Schaeffer P, Hirschbein L (1982) Phenotypic expression in vivo and transforming activity in vitro: Two related functions of folded bacterial chromosomes. Mol Gen Genet 185:65–68Google Scholar
- Ferenczy L (1981) Microbial protoplast fusion. In: Glover SW, Hopwood DA (eds) Genetics as a tool in microbiology. Cambridge University Press, p 1–34Google Scholar
- Fodor K, Alfoldi L (1976) Fusion of protoplasts of Bacillus megaterium. Proc Natl Acad Sci USA 73:2147–2150Google Scholar
- Guillén N, Le Hegarat F, Fleury AM, Hirschbein L (1978) Folded chromosome of vegetative Bacillus subtilis: Composition and properties. Nucl Acids Res 5:475–490Google Scholar
- Guillén N, Gabor MH, Hotchkiss RD, Hirschbein L (1982) Isolation and characterization of the nucleoid of non-complementing diploids from protoplast fusion in Bacillus subtilis. Mol Gen Genet 185:69–74Google Scholar
- Hirschbein L, Guillén N (1982) Characterization, assay and use of isolated bacterial nucleoids. In: Methods of biochemical analysis, vol 28. John Wiley and Sons, New York, pp 297–327Google Scholar
- Hotchkiss RD, Gabor MH (1980) Biparental products of bacterial protoplast fusion showing unequal parental chromosome expression. Proc Natl Acad Sci USA 77:3553–3557Google Scholar
- Isaacs LN, Echols H, Sly WS (1965) Control of lambda mRNA by the cI immunity region. J Mol Biol 13:963–967Google Scholar
- Kao K, Michayluk MR (1974) A method for high frequency intergenic fusion of plant protoplasts. Planta 115:355–367Google Scholar
- Le Hegarat F, Guillen N, Fleury AM, Hirschbein L (1978) Comparative studies on purified nucleoids of Bacillus subtilis during growth and sporulation. In: Chambliss G, Vary JC (eds) Spores VII. American Society for Microbiology, Washington DC, pp 193–201Google Scholar
- Lévi-Meyrueis C, Sanchez-Rivas C, Schaeffer p (1980) Formation de bactéries diploides stables par fusion de protoplastes de Bacillus subtilis et effet de mutations rec sur les produits de fusion formés. CR Acad Sci Paris D 291:67–70Google Scholar
- Maizels N (1976) Dictyostelium 17S, 25S, and 5S rDNAs lie within a 38,000 base pair repeated unit. Cell 9:431–438Google Scholar
- Rutberg L (1969) Mapping of a temperate bacteriophage active on Bacillus subtilis. J Virol 3:38–44Google Scholar
- Rutberg L (1973) Heat induction of prophage ϕ105 in Bacillus subtilis: bacteriophage induced bidirectional replication of the bacterial chromosome. J Virol 12:9–12Google Scholar
- Rutberg L, Sandstrom E, Nilson K (1968) Circular chromosomal map of a temperate Bacillus phage. Virology 32:103–108Google Scholar
- Sanchez-Rivas C (1982) Direct selection of complementing diploids from PEG-induced fusion of Bacillus subtilis protoplasts. Mol Gen Genet 185:329–333Google Scholar
- Sanchez-Rivas C, Garro A (1979) Bacterial fusion assayed by a prophage complementation test. J Bacteriol fusion assayed by a prophage complementation test. J Bacteriol 137:1340–1345Google Scholar
- Sanchez-Rivas C, Lévi-Meyrueis C, Lazard-Monier F, Schaeffer P (1982) Diploid state of phenotypically recombinant progeny arising after protoplast fusion in Bacillus subtilis. Mol Gen Genet 188:272–278Google Scholar
- Schaeffer P, Ionesco H, Ryter A, Balassa G (1963) La sporulation de Bacillus subtilis: étude génétique et physiologique. Coll Int CNRS 124:553–563Google Scholar
- Schaeffer P, Hotchkiss RD (1978) Fusion of bacterial protoplasts. Methods Cell Biol 20:149–158Google Scholar
- Schaeffer P, Cami B, Hotchkiss RD (1976) Fusion of bacterial protoplasts. Proc Natl Acad Sci USA 77:2151–2155Google Scholar
- Szybalski W, Nyer V (1967) The mitomycin and porfiromycin. In: Gottlieb D, Shaw P (eds) Antibiotics, vol I. Springer Berlin Heidelberg New York, pp 211–244Google Scholar
- Witkin E (1976) UV mutagenesis and inducible DNA repair in Escherichia coli. Bacteriol Rev 40:869–907Google Scholar
- Williams JG, Lloyd MM, Devine JM (1979) Characterization and transcription analysis of a cloned sequence derived from a major developmentally regulated mRNA of D. discoideum. Cell 17:903–913Google Scholar
- Yamamoto KR, Alberts M, Benzinger R, Lawhorne L, Treiber G (1970) Rapid bacteriophage sedimentation in the presence of polyethylene glycol and its application to large scale virus purification. Virology 40:734–744Google Scholar
- Yasbin RE (1977) DNA repair in B. subtilis. I. The presence of an inductible system. Mol Gen Genet 153:211–218Google Scholar